Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.

 
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
Australian
 Biochemist
 The Magazine of the Australian
 Society for Biochemistry and
 Molecular Biology Inc.
 December 2020, Volume 51, Number 3

 ISSN 1443-0193
VOL 51 NO 3 DECEMBER 2020   AUSTRALIAN BIOCHEMIST   PAGE 1
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
Table of Contents
  3      Editorial Committee
  4      From the President
  5      ASBMB 2020 Meeting Reports
  9      Publications with Impact
         Unravelling the Speciation, Trafficking, Autoinflammatory Potential of the Killer Protein, MLKL
         Fluid Coupling in the Rotary Motor of Life
         The Natural Function of a Multidrug Resistance Transporter
         Uncovering a New Way in Which Tumours Co-opt Their Microenvironments to Promote
         Disease Progression
         Mutational Landscape of Gall Bladder Cancers
         Structural Venomics: from Single to Double Knots and Everything in Between
  19     ASBMB Education Feature
         Online Lessons From Our Students – 2020 ASBMB Education Symposium
         Ten Reasons to Crowdsource Exam Questions (and How to Do it Properly)
         Transitioning Case-based Biochemistry Workshops to Online During COVID-19
  25     SDS Page
         A Beginner’s Guide to Milestone Preparation
  27     Competition: Campus Visit
  28     Perth Protein Group: an ASBMB Special Interest Group
  29     Metabolism and Molecular Medicine: an ASBMB Special Interest Group
  31     Off the Beaten Track
         Follow Your Dreams, Until Something Better Comes Along
  32     ASBMB Member Wins Western Australian Premier’s Science Award
  34     Intellectual Property
         Timing is Everything: When to Pull the Trigger on Patent Filing
  37     News from the States
  41     Great Expectations
         It’s Not Only What You Know, It’s Who                                                Front Cover
         You Know – an Unplanned Career
                                                                            ASBMB President, Joel Mackay,
  45     ASBMB Member Named Melburnian                             delivers the opening address at the 2020
         of the Year                                              ASBMB Research Symposium held online.
  45     In Memoriam
  47     ASBMB Annual Reports
  51     Our Sustaining Members                                                      The Australian Biochemist
                                                                              Editor Tatiana Soares da Costa
  56     ASBMB Council                                                       Editorial Officer Liana Friedman
  57     Directory                                                  © 2020 Australian Society for Biochemistry
                                                                  and Molecular Biology Inc. All rights reserved.

PAGE 2                                    AUSTRALIAN BIOCHEMIST                       VOL 51 NO 3 DECEMBER 2020
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
Australian Biochemist Editorial Committee
                        Editor                                        Editorial Officer
                        Dr Tatiana Soares da Costa                    Liana Friedman
                        Department of Biochemistry and                Email: liana.friedman@monash.edu
                        Genetics
                        La Trobe Institute for Molecular
                        Science
                        La Trobe University
                        Bundoora VIC 3086
                        Email: editor@asbmb.org.au
                        Phone: (03) 9479 2227

                        Dr Doug Fairlie                               Dr Sarah Hennebry
                        Olivia Newton-John Cancer                     FPA Patent Attorneys
                        Research Institute and La Trobe               101 Collins Street
                        University                                    Melbourne VIC 3000
                        Heidelberg VIC 3084                           Email: sarah.hennebry@
                        Email: doug.fairlie@onjcri.org.au             fpapatents.com
                        Phone: (03) 9496 9369                         Phone: (03) 9288 1213

                        Joe Kaczmarski                                 Associate Professor Tracey Kuit
                        Research School of Chemistry                   School of Chemistry and Molecular
                        Australian National University                 Bioscience
                        Canberra ACT 0200                              University of Wollongong
                        Email: joe.kaczmarski@                         Wollongong NSW 2522
                        anu.edu.au                                     Email: tracey_kuit@uow.edu.au
                                                                       Phone: (02) 4221 4916

                        Dr Erinna Lee                                  Dr Nirma Samarawickrema
                        La Trobe Institute for Molecular               Department of Biochemistry and
                        Science and Olivia Newton-John                 Molecular Biology
                        Cancer Research Institute                      Monash University
                        Heidelberg VIC 3084                            Clayton VIC 3800
                        Email: erinna.lee@latrobe.edu.au               Email: nirma.samarawickrema@
                        Phone: (03) 9496 9369                          monash.edu
                                                                       Phone: (03) 9902 0295

                        Dr Gabrielle Watson
                        Monash Biomedicine Discovery
                        Institute
                        Monash University
                        Clayton VIC 3800
                        Email: gabrielle.watson@
                        monash.edu
                        Phone: (03) 9902 9227

VOL 51 NO 3 DECEMBER 2020                     AUSTRALIAN BIOCHEMIST                             PAGE 3
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
From the President
  This piece is my swan song as ASBMB President, as               One of our aims this
I hand over the reins to Jacqui Matthews for the next           year was to increase our
two years. Jacqui will be well known to many of you             involvement in science
in the Australian biochemistry and molecular biology            advocacy. We achieved
community, particularly those of you who attend the             an important first step in
Lorne Conference on Protein Structure and Function.             that direction – Tatiana
Jacqui has just finished up her term as Chair of that           Soares da Costa has
meeting and was obviously looking for a new challenge!          recently been elected to
  The last two years have seen some change at the               the board of Science and
ASBMB, particularly in our trialling of a new Society           Technology         Australia
conference timetable. We decided to host an ASBMB-              (STA) – the advocacy
focussed meeting every two years and have ComBio                group that represents
only in the alternate years. We thought this might offer        over      80,000      STEM
members the opportunity to meet in a more intimate              professionals, including
venue than one of the large convention centres that we          ASBMB’s members, to government – as the Biological
need to use for ComBio meetings.                                Sciences Cluster Representative. We look forward to
  Our first ASBMB-only meeting was held in Perth in             hearing from her about the role and about what we
October 2019. Nic Taylor from the University of Western         can do to further the cause of science on the political
Australia put a huge effort into coordinating a team to         agenda in Australia.
run this meeting. The overall consensus amongst the               What else have I been trying to do during my term as
attendees was that it was a very enjoyable meeting with         President? We have done quite a bit of ‘internal’ work,
a good atmosphere and a great program. Attendance               clarifying guidelines for State Representatives and
at the meeting was lower than we would have hoped               Special Interest Groups, providing State Representatives
for, no doubt in part due to it being situated on the           with a forum to connect with each other on a regular
west coast, and possibly also partly due to uncertainty         basis to exchange ideas, and reinvigorating the website
on our part about the best way to focus the topics of           (thanks to our webmaster Liana Friedman!). I would
the meeting. When we were planning the meeting, we              still like to survey members about the Society and to
thought that highlighting a small number of topics in the       consider whether we should coordinate an annual
program (while still allowing talks in any subject area)        meeting of Heads of Discipline in our field, as a way to
might attract more attendees. With hindsight, we may            share knowledge. I am hoping to organise these things
have been wiser to not showcase particular subject              before the end of 2020.
areas, as it might have given the impression that other           I’d like to finish off by giving enormous thanks to the
topics would not be covered at all during the meeting.          people who have contributed to ASBMB during my time
  We are still keen to see if we can get the ASBMB-only         as President and who, without exception, have been
format to work – scientifically and financially – and I think   inspired, efficient, thorough and good humoured. Briony
we will try to cast the net wider in future incarnations of     Forbes and Dominic Ng have been stalwart Secretaries,
the meeting. We will also seek feedback from members            Marc Kvansakul has been a trustworthy Treasurer,
to gauge their thoughts on meeting formats.                     Tatiana Soares da Costa, an energetic Editor (and
  Speaking of meetings, the online ASBMB symposia               savvy social media specialist – follow @ITSASBMB on
in September were a real success and it makes me                Twitter), Liana Friedman, our expert Webmaster and
wonder whether we could continue the momentum                   Editorial Officer, and Leann Tilley as Past President.
with that format of having one or more short online             And of course, Sally and Chris Jay have been the
meetings (or even mixed live plus online, once that’s           consummate professionals who have continued to hold
permitted) as part of our regular offering (alongside           it all together for us at the National Office – and have
face-to-face conferences). I know most of us prefer the         engaged with our ever-valuable Sustaining Members.
face-to-face format, but there are some advantages in           Jackie Wilce and Mark Hulett have allowed me to not
terms of ease of organisation, low cost of attendance           be concerned at all about the next ComBio, by doing
and the opportunity to give more members a chance to            a superb job of organising the one that will be held in
present their work (particularly ECRs). The Education-          Melbourne in 2022 (and locking in the 2020 Nobel Prize
focussed meeting was particularly well attended, and            in Chemistry Jennifer Doudna as a keynote speaker).
I really would like to see us take advantage of that            Leann has also strengthened our conference portfolio
success to create a regular forum for presentations             by leading a team to run the IUBMB Congress in
and discussions in the biochemical education space.             Melbourne in 2024. Having people doing great jobs like
Encouraging the Society to increase its focus on                this makes the role of the President infinitely easier!
biochemistry education has been on my agenda, and                 Finally, thank you all too as members – you of course
I am pleased that we have attracted an Education                are the Society. See you all at the next meeting!
Plenary speaker for ComBio2022.                                                                              Joel Mackay
                                                                                                       President, ASBMB
PAGE 4                                           AUSTRALIAN BIOCHEMIST                        VOL 51 NO 3 DECEMBER 2020
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
ASBMB 2020 Meeting
With the postponement of ComBio2020 until 2022, ASBMB
hosted a virtual meeting in 2020. The ASBMB Council felt it
was important to provide our members with opportunities to
come together to maintain a sense community and celebrate
our award winners. We held an Education Symposium chaired
by Nirma Samarawickrema on 29 September and a Research
Symposium chaired by Tatiana Soares da Costa on 30 September.

Education Symposium
  COVID-19 was the thunderbolt that hit our universities      they experienced as a result of the sudden shift to remote
in 2020 and forced educators into remote teaching.            learning, the strategies they used to adapt, and their
Biochemistry educators, like other educators worldwide,       visions for a post-COVID learning environment. These
scrambled behind the scenes, urgently transforming their      presentations were timely reminders to educators of the
teaching for online delivery while ensuring continuity,       criticality of our ultimate audience – our students.
quality and integrity. This unexpected and rapid transition
to online remote learning resulted in the reinvention of
much of our teaching and learning, and the adoption
of a wealth of strategies and novel practices to engage
students in workshops, tutorials, practical classes and
assessment.
  The ASBMB Education Symposium – Teaching
Remotely, Sharing Practice was held on 29 September
2020. The Education Symposium provided national
and international participants a platform to share their
insights and experiences, innovations and good practice       Biochemistry students Ben Urzua and Rhianna Coscia.
to transform the student learning experience as we move
forward.                                                        The symposium had 250 registrants, with 131 from
  This event offered a mix of short and extended              across Australia representing all six states and a territory
presentations from educators who demonstrated how             and 119 from overseas, many of whom were members of
they:                                                         the Federation of Asian and Oceanian Biochemists and
• Motivated students to learn online through game-            Molecular Biologists (FAOBMB).
    based platforms, digital portfolios, Zoom case-study        ASBMB President, Professor Joel Mackay, attributed
    workshops.                                                the great level of participation in our event to the mutual
• Transformed large face-to-face lab classes to online        commitment and drive of biochemistry educators to
    laboratory classes without losing the interactivity       enhance teaching practices and the value they place
    of the face-to-face classes by using a variety of         on educating and inspiring our future generations of
    strategies including Zoom to guide a metabolomics         scientists. This sentiment was echoed by Professor
    computer practical, PYMOL and Sketchfab to map            Roger Daly (Head, Biochemistry and Molecular Biology,
    out and create guided tours of a protein.                 Monash University) in his welcome address.
• Developed skills such as evaluative judgement,                The symposium showcased a snapshot of the innovations
    research and writing skills through online                implemented in remote learning. It is now up to the broader
    approaches.                                               education community to reflect upon, adapt and apply this
• Created authentic assessments even for large-scale          learning as it is relevant to our specific student cohorts in
    undergraduate research experiences, and created           a post-COVID world. Professor Liz Johnson (Deputy Vice-
    online assessments to avoid plagiarism through the        Chancellor, Education, Deakin University) endorsed this
    development of software that could generate unique        idea in her keynote address, Online is the New Normal:
    data sets.                                                Teaching as We Work. “Disruption creates opportunity as
  Contributions from our students were a highlight of the     assumptions and conventions are challenged. It is time
conference. Third year student Benjamin Urzua (Monash         to re-examine the value of each teaching mode and to
University) and second year student Rhianna Coscia            select learning and assessment activities that leverage
(University of South Australia) discussed the challenges      the strengths of online as well as face-to-face teaching.”

VOL 51 NO 3 DECEMBER 2020                      AUSTRALIAN BIOCHEMIST                                               PAGE 5
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
ASBMB 2020 Meeting

Keynote speaker Liz Johnson.                            Sophie Paquet-Fifield.

  We congratulate the Education Symposium winners:        We thank the ASBMB for supporting the vision of this
Best abstract (sponsored by Fisher Biotec Australia)    inaugural online symposium, Fisher Biotec for sponsoring
Professor Gareth Denyer and Dr Alice Huang              the best abstract award and Danielle Rika from the
University of Sydney                                    Department of Biochemistry and Molecular Biology,
Generation of unique datasets to complement online      Monash University, for her outstanding support to the
practicals and build student experimental design and    Education Symposium Committee and her meticulous
troubleshooting skills                                  attention to detail. As the ASBMB Education Symposium
                                                        Organising Committee, we gratefully acknowledge each
                                                        other and unanimously agree that we were a wonderful
                                                        team!
                                                          Our sincere thanks to our student participants, our
                                                        presenters who shared their practice and to all who
                                                        attended. The presentations highlight how educators rose
                                                        to the challenge of remote teaching.
                                                          Videos of the day’s presentations are available online.

                                                                             ASBMB Education Symposium
                                                                                      Organising Committee
Gareth Denyer and Alice Huang.                                  Nirma Samarawickrema, Monash University
                                                                      Tracey Kuit, University of Wollongong
Highly Commended Abstract (sponsored by ASBMB)              Amber Willems-Jones, University of Melbourne,
Dr Kathryn Jones and Dr Monica Kan                                  Matthew Clemson, University of Sydney
University of Auckland, New Zealand                         Maurizio Costabile, University of South Australia
Creating interactive online biochemistry laboratories
using H5P

Kathryn Jones and Monica Kan.
Best Zoom Background (sponsored by ASBMB)
Dr Sophie Paquet-Fifield
University of Melbourne
For a background created collaboratively by her young
children and students

PAGE 6                                       AUSTRALIAN BIOCHEMIST                    VOL 51 NO 3 DECEMBER 2020
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
ASBMB 2020 Meeting
Research Symposium
  On 30 September, we held a Research Symposium,
chaired by Tatiana Soares da Costa (ASBMB Editor
and Chair of Communications) with the help of State
Representatives: Benjamin Schulz, Erinna Lee, Kate
Brettingham-Moore, Kate Quinlan, Melissa Pitman and
Monika Murcha.
  We had an exciting Research Symposium program
with two engaging plenary speakers. Professor Shelley
Berger (University of Pennsylvania) started her talk
discussing how epigenetic changes and pathways govern
social behaviour in ants and ended with how chromatin
enzymes modify the tumour suppressor p53. Professor         National University) provided a great overview of the
Glenn King (University of Queensland) gave a fascinating    protein work his lab does, with a focus on the evolution
talk on how a spider venom peptide can be used to treat     of binding specificity in the amino acid binding proteins.
ischemic injuries of the heart and brain.                   Eppendorf Edman ECR Award winner Professor Si Ming
                                                            Man (Australian National University) discussed the role of
                                                            GBPs in inflammasome activation. We also had lightening
                                                            talks from ASBMB Travel Fellowship awardees, Dr Amy
                                                            Baxter (La Trobe University), Dr Steffi Cheung (University
                                                            of Melbourne), Dr Mengjie Hu (University of Melbourne)
                                                            and Anukriti Mathur (Australian National University).
                                                              The ASBMB AGM was held after lunch and with many of
                                                            our members attending, we had no issues with reaching
                                                            a quorum! Associate Professor Dominic Ng from the
                                                            University of Queensland was elected as ASBMB National
                                                            Secretary, taking over from Professor Briony Forbes,
                                                            who we thanked for her many years of hard work and
                                                            stewardship of the Society. The changing of the guard
                                                            continued as we welcomed new State Representatives
                                                            for ACT, NSW, QLD, TAS and VIC. At the conclusion of
                                                            the symposium, we had a happy hour using Spatial Chat,
                                                            which allowed us to ‘move around’ a crowd to join and
                                                            leave conversations, replicating as much as possible the
                                                            conference cocktail mixers we have sorely missed.
                                                              Research Symposium attendance numbers were
                                                            pleasing, with over 260 registrants from around Australia
                                                            and overseas. The two-day ASBMB meeting highlighted
                                                            the amazing advances our Australian biochemistry and
                                                            molecular biology community is making in education
                                                            and research. Thanks to the Organising Committee, the
Presentations by plenary speakers Shelley Berger and        award sponsors and ASBMB President, Joel Mackay, for
Glenn King.                                                 their support.
                                                              Videos of the day’s presentations are available online.
 We also heard from our 2020 ASBMB awardees.                  Planning continues for 2021, with an expanded East
Lemberg medallist Professor Trevor Lithgow (Monash          Coast Protein Meeting (more details in the coming
University) described how porin loss leads to carbapenem    months) and the FAOBMB Congress in Christchurch,
resistance in Klebsiella. SDR Scientific Education Award    New Zealand. We look forward to seeing you then –
winner Dr Nirma Samarawickrema (Monash University)          hopefully face-to-face!
presented recommendations for education amidst COVID                                        Tatiana Soares da Costa
based on case studies. Boomerang Award winner Dr                         Chair, ASBMB 2020 Research Symposium
Matthew Doyle (National Institutes of Health Bethesda)                     La Trobe Institute for Molecular Science
discussed how BamA forms a translocation channel for                                              La Trobe University
secretion of bacterial autotransporter proteins. Shimadzu                         T.SoaresdaCosta@latrobe.edu.au
Research medallist Professor Colin Jackson (Australian                                   Twitter: @Tatiana_Biochem

VOL 51 NO 3 DECEMBER 2020                     AUSTRALIAN BIOCHEMIST                                            PAGE 7
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
PAGE 8   AUSTRALIAN BIOCHEMIST   VOL 51 NO 3 DECEMBER 2020
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
Publications with Impact
      Publications with Impact profiles recent, high impact publications by ASBMB members.
           These short summaries showcase some of the latest research by presenting
             the work in a brief but accessible manner. If your work has recently been
               published in a high profile journal, please email editor@asbmb.org.au.

        Unravelling the Speciation, Trafficking,
Autoinflammatory Potential of the Killer Protein, MLKL
 Our team at the Walter and Eliza Hall Institute of Medical Research has published a trio of studies in
Nature Communications that shed new light on the protein, MLKL, and how it causes an inflammatory form
of cell death known as necroptosis. As outlined below, these studies use structural biology to understand
evolutionary changes in MLKL, advanced microscopy to visualise when and where MLKL is activated in
cells undergoing necroptosis, and genetic models to address how mutations in MLKL can trigger lethal
auto-inflammatory disease.

      Davies KA, Fitzgibbon C, Young SN, Garnish SE, Yeung W, Coursier D, Birkinshaw RW,
   Sandow JJ, Lehmann WIL, Liang LY, Lucet IS, Chalmers JD, Patrick WM, Kannan N, Petrie EJ,
   Czabotar PE*, Murphy JM*. Distinct pseudokinase domain conformations underlie divergent
     activation mechanisms among vertebrate MLKL orthologues. Nat Commun 2020;11:3060.
              *Corresponding authors: czabotar@wehi.edu.au, jamesm@wehi.edu.au

  As    a    pseudokinase,
MLKL is unable to catalyse
phosphotransfer reactions.
Instead, phosphorylation
of     MLKL’s      activation
loop by the upstream
kinase, RIPK3, is thought
to toggle a molecular
switch, such that the
killer N-terminal domain
becomes exposed and can
permeabilise the plasma
membrane to induce cell
death by necroptosis. The Crystal structures of the rat and horse MLKL pseudokinase domain, that were used to
underlying      mechanism better understand species compatibility. PDB: 6VBZ, 6VC0.
was recently shown to
differ between mouse and human necroptosis, which between MLKL orthologues where they could seldom
led us to examine the extent of divergence among ‘talk’ to RIPK3 within mouse and human cells. Together,
vertebrate MLKL orthologues. By studying nine MLKL our findings suggest that the MLKL:RIPK3 cassette has
orthologues spanning multiple phylogenies, and solving rapidly co-evolved in different species, potentially due
the structures of the rat and horse MLKL pseudokinase to varying selection pressures exerted by the pathogens
domains, we identified a profound lack of compatibility that target each organism.

                                                        From left: André Samson,
                                                        Joanne Hildebrand and
                                                        Katherine Davies.

VOL 51 NO 3 DECEMBER 2020                   AUSTRALIAN BIOCHEMIST                                       PAGE 9
Australian Biochemist - The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc.
Publications with Impact
  Samson AL*, Zhang Y, Geoghegan ND, Gavin XJ, Davies KA, Mlodzianoski MJ, Whitehead LW,
 Frank D, Garnish SE, Fitzgibbon C, Hempel A, Young SN, Jacobsen AV, Cawthorne W, Petrie EJ,
       Faux MC, Shield-Artin K, Lalaoui N, Hildebrand JM, Silke J, Rogers KL, Lessene G,
     Hawkins ED*, Murphy JM*. MLKL trafficking and accumulation at the plasma membrane
         control the kinetics and threshold for necroptosis. Nat Commun 2020;11:3151.
*Corresponding authors: samson.a@wehi.edu.au, hawkins.e@wehi.edu.au, jamesm@wehi.edu.au

  MLKL normally resides in the cytoplasm, but during
necroptosis it translocates to membranes and kills cells
by disrupting plasma membrane integrity. In this study,
we used single-cell imaging to meticulously map the
activation and relocation of endogenous human MLKL
during necroptosis. We found that, once activated, MLKL
moves to the plasma membrane via Golgi-, tubulin- and
actin-dependent mechanisms, and that inhibiting these
trafficking mechanisms specifically slows the onset of
necroptotic cell death. Strikingly, during necroptosis,
MLKL co-trafficks with tight junction proteins – proteins
best known for their role in epithelial barrier formation.
This co-trafficking allows MLKL to accumulate into
micron-sized ‘hotpots’ at the plasma membrane during
necroptosis. In this work, we identified two new rate-
limiting checkpoints in necroptosis: (1) a trafficking       Two cells undergoing necroptosis exemplifying the
checkpoint that controls the movement of MLKL to             junctional accumulation of human MLKL.
the cell periphery, and (2) a junctional checkpoint
that renders necroptosis ‘contagious’ and dictates the
amount of MLKL needed at the plasma membrane to
trigger cell death.

        Hildebrand JM*, Kauppi M, Majewski IJ, Liu Z, Cox AJ, Miyake S, et al., Murphy JM,
        Alexander WS*, Silke J*. A missense mutation in the MLKL brace region promotes
    lethal neonatal inflammation and hematopoietic dysfunction. Nat Commun 2020;11:3150.
 *Corresponding authors: jhildebrand@wehi.edu.au, alexandw@wehi.edu.au, silke@wehi.edu.au
 The signalling pathway that culminates in MLKL              MLKL point mutant mouse strain (MlklD139V) in an ENU
activation and killer activity has many entry and branch     mutagenesis screen long before MLKL was implicated
points, making it tough to tease apart the precise role of   in necroptosis, where homozygous pups died soon after
MLKL-mediated cell lysis in the aetiology of inflammation    birth. Our team found that this point mutation conferred
and disease. Serendipitously, our team identified a          both constitutive activity to MLKL at the molecular level,
                                                             but that this deadly activity was very efficiently ‘kept in
                                                             check’ at the cellular level through MLKL proteolysis
                                                             below a threshold. This threshold is overwhelmed only
                                                             when two MlklD139V alleles are inherited, and only after
                                                             birth – physiologically manifesting as inflammation in/
                                                             around the lower head and mediastinum (salivary
                                                             glands, heart and thymus). We are excited to continue
                                                             exploring the inflammatory role of a very closely situated
                                                             cluster of human MLKL mutations which are present in
An activating mutation in mouse MLKL manifests in a          up to 8% of individuals globally, and enriched (in trans)
deadly postnatal inflammatory syndrome. Longitudinal         in a form of paediatric bone disease, CRMO.
section of 2-day old mouse mediastinum.

                                                                                Katherine Davies, André Samson,
                                                                           Joanne Hildebrand and James Murphy
                                                               Walter and Eliza Hall Institute of Medical Research
                                                             Department of Medical Biology, University of Melbourne

PAGE 10                                        AUSTRALIAN BIOCHEMIST                        VOL 51 NO 3 DECEMBER 2020
Publications with Impact
             Fluid Coupling in the Rotary Motor of Life
   Sobti M, Walshe JL, Wu D, Ishmukhametov R, Zeng YC, Robinson CV, Berry RM, Stewart AG*.
              Cryo-EM structures provide insight into how E. coli F1Fo ATP synthase
                accommodates symmetry mismatch. Nat Commun 2020;11:2615.
                     *Corresponding author: a.stewart@victorchang.edu.au
  F1Fo ATP synthase is a complex macromolecular
machine consisting of two coupled rotary motors:
the soluble F1 ATPase and the transmembrane
Fo motor. The F1 ATPase is a chemical motor
that synthesises adenosine triphosphate (ATP),
whereas the Fo motor is an electric motor powered
by proton flow. Together these motors make a major
contribution to the generation of cellular chemical
energy by converting the proton motive force (pmf)
into ATP, the universal cellular energy currency.
By imaging the intact enzyme from Escherichia
coli using cryo-electron microscopy (cryo-EM), the
Stewart lab and their collaborators were able to
generate detailed atomic-level information on how
energy is transferred elastically between the two            The Stewart laboratory, from left: James Walshe, Yi Zeng,
motors.                                                      Alastair Stewart and Meghna Sobti.
  Recent improvements in cryo-EM have produced
new insight on F1Fo ATP synthase, providing detailed         resides in the membrane and is comprised of a ring of
information about how this enzyme functions at the           c-subunits (the c-ring) that can rotate relative to a stator,
atomic level. Bacterial F1Fo ATP synthase has been           the a-subunit. The motor is driven by the pmf, generated
studied for many decades and has been used as a model        by oxidative phosphorylation or photophosphorylation,
system to understand ATP synthase function. The F1           and acts like a horizontal water wheel, with protons
ATPase is comprised of a hexameric ring of alternating       sequentially binding to the c-ring from the periplasm
a- and b-subunits, with a single g subunit bound             and rotating through it before being released into the
asymmetrically in the ring centre. As this central ‘rotor’   cytoplasm. The rotation of the c-ring is transferred to the
g-subunit rotates, it generates conformational changes       central rotor (g subunit), thereby coupling the rotation of
in the a- and b- subunits. The F1 ATPase enzyme              one motor to the other. An interesting feature of almost
operates with a binding-change mechanism, whereby            all ATP synthases is a symmetry mismatch between the
rotation of the central rotor induces sequential binding     F1 and Fo motors because the F1 ATPase has threefold
of ADP and inorganic phosphate in the b-subunits,            rotational symmetry, whereas the Fo motor has tenfold
which are then combined to make ATP. The Fo motor            symmetry (in E. coli). This mismatch causes a non-
                                                                                           The cryo-EM structure of
                                                                                           E. coli F1Fo ATP synthase.
                                                                                           a. Sorting of the particles
                                                                                              using 3D classification
                                                                                              resulted in nine discrete
                                                                                              sub-states.
                                                                                           b. The sub-states show
                                                                                              rotational ‘sub-stepping’
                                                                                              in the Fo motor, which is
                                                                                              mediated by a flexible
                                                                                              peripheral stalk.
                                                                                           c. Local resolution of
                                                                                              the F1 ATPase and
                                                                                              composition of the
                                                                                              catalytic sites.
                                                                                           d. Lipids (wheat density)
                                                                                              bridge protein subunits
                                                                                              in the Fo motor.

VOL 51 NO 3 DECEMBER 2020                      AUSTRALIAN BIOCHEMIST                                             PAGE 11
Publications with Impact
integral H+/ATP ratio and consequently, the motors need     store elastic energy in the ‘peripheral’ stalk to overcome
to be coupled elastically to operate efficiently.           the symmetry mismatch between the two motors (b).
  High resolution cryo-EM maps of the intact E. coli F1Fo   High resolution information in the catalytic domain also
ATP synthase were generated by imaging the detergent-       gave insight into how the F1 ATPase is inhibited from
solubilised complex in the presence of MgADP. These         operating in reverse (c), and maps focussing on just the
maps highlighted nine different sub-states that described   Fo motor showed a novel lipid interaction that can bridge
the molecular motion of the complex, nucleotide binding     a subunit within the c-ring with the stator a subunit (d),
occupancy, and lipid-mediated interactions (see figure).    thereby increasing the interaction surface which may
Of greatest interest was the ‘sub-stepping’ observed in     aid fluid coupling.
the Fo motor, which showed how the transmembrane                                                      Alastair Stewart
motor can rotate independently of the F1 ATPase and                       Victor Chang Cardiac Research Institute

                         The Natural Function of a
                      Multidrug Resistance Transporter
          Shafik SH, Cobbold SA, Barkat K, Richards SN, Lancaster NS, Llinás M, Hogg SJ,
            Summers RL, McConville MJ, Martin RE*. The natural function of the malaria
             parasite’s chloroquine resistance transporter. Nat Commun 2020;11:3922.
                        *Corresponding author: rowena.martin@anu.edu.au

 “What is the natural function of the Plasmodium            as a drug target. In this study, the Martin lab and
falciparum chloroquine resistance transporter               collaborators at the University of Melbourne and
(PfCRT)?” has been a major unanswered question              the Pennsylvania State University provide the long-
in malaria parasite biology for two decades.                awaited elucidation of PfCRT’s native substrates
Mutations in PfCRT were originally identified as            and normal physiological role. PfCRT functions
the main cause of chloroquine resistance in P.              to export host-derived peptides containing four to
falciparum, but are now known to play a key role            eleven residues from the digestive vacuole to the
in multidrug resistance. The transporter resides at         cytosol, thereby providing a source of amino acids
the membrane of the parasite’s digestive vacuole            for parasite metabolism and preventing the osmotic
– a lysosomal-type organelle in which many                  stress of this organelle.
antimalarial drugs accumulate, act, and/or are               The malaria parasite consumes vast quantities of host
activated. Mutant isoforms of PfCRT contribute              proteins (mainly haemoglobin) to provide the space and
to multidrug resistance primarily by transporting           amino acids it requires for proliferation within the host
drugs out the vacuole. PfCRT is also essential              erythrocyte, and also to maintain the osmotic balance
for parasite survival and has itself been identified        of the parasitised cell. This process occurs within the

                                                                                             Wildtype (wt) PfCRT
                                                                                             exports peptides from
                                                                                             the digestive vacuole
                                                                                             into the parasite cytosol.
                                                                                             The diminished capacity
                                                                                             of the drug resistance
                                                                                             conferring isoform (mut)
                                                                                             for peptide transport
                                                                                             causes a build-up of
                                                                                             peptides and amino
                                                                                             acids, thereby raising
                                                                                             the vacuole’s osmotic
                                                                                             pressure and reducing
                                                                                             the rate of digestion.

PAGE 12                                       AUSTRALIAN BIOCHEMIST                       VOL 51 NO 3 DECEMBER 2020
Publications with Impact
                                                            eleven residues, varying in both charge and composition,
                                                            are substrates of PfCRT in vitro and in situ. Furthermore,
                                                            we showed that the protein does not transport other
                                                            metabolites and/or organic or inorganic ions. The
                                                            transport of peptides and peptide mimics via PfCRT is
                                                            saturable, can be blocked by known PfCRT inhibitors
                                              Sarah         (e.g. verapamil), and is dependent on protons as well as
                                              Shafik        on a second solute that remains to be identified, but which
                                              (left) and    is naturally present in the Xenopus oocyte. Relative to
                                              Rowena        wildtype PfCRT, the drug resistance conferring isoforms
                                              Martin.       exhibit reduced rates of peptide transport and accept
                                                            a narrower range of peptides and peptide mimics. The
acidic environment of the parasite’s digestive vacuole.     reduced capacity of mutant PfCRT to efflux peptides
Importantly, alterations to PfCRT can affect the normal     out of the vacuole accounts for the accumulation of
physiology of this organelle as well as the growth of the   these substrate peptides in the drug resistant lines, and
parasite. For example, downregulation of PfCRT causes       explains the impaired fitness of parasites expressing
the vacuole to swell and impairs parasite growth, and       mutant isoforms of the transporter. The physiological
potent inhibition of mutant PfCRT results in parasite       relevance of this correlation was further substantiated
death. Moreover, the mutant isoforms that confer drug       when we used the peptidomic datasets to correctly
resistance typically impose a fitness cost upon the         predict host-derived peptides as being substrates or
parasite, the severity of which is dependent upon the       non-substrates of PfCRT. Our work indicates that the
number and nature of the mutations.                         PfCRT-mediated transport of peptides from the vacuole
  Attempts to use heterologous expression systems to        to the cytosol serves to (1) provide a source of amino
identify the native substrates of PfCRT in vitro have       acids to support the parasite’s high growth rate, and (2)
produced wildly different ‘natural functions’ for the       reduce peptide levels within the vacuole and thereby
protein, with there being no convincing support for         prevent the osmotic stress, swelling, and dysfunction
one claim over the others. We employed the Xenopus          of this organelle. The resolution of PfCRT’s native
oocyte expression system, as well as measurements           substrate-specificity and physiological role provides a
of solute transport, drug activity, and metabolite levels   foundation for the development of drugs that target both
in isogenic parasite lines expressing different pfcrt       its normal and drug resistance conferring functions.
isoforms, to identify, and independently validate, the                                                Rowena Martin
natural substrates of PfCRT.                                                           Research School of Biology
  Our work revealed that host-derived peptides of four to                            Australian National University

Uncovering a New Way in Which Tumours Co-opt Their
 Microenvironments to Promote Disease Progression
     Boyle ST, Poltavets V, Kular J, Pyne NT, Sandow JJ, Lewis AC, Murphy KJ, Kolesnikoff N,
    Moretti PAB, Tea MN, Tergaonkar V, Timpson P, Pitson SM, Webb AI, Whitfield RJ, Lopez AF,
       Kochetkova M*, Samuel MS*. ROCK-mediated selective activation of PERK signalling
               causes fibroblast reprogramming and tumour progression through a
                CRELD2-dependent mechanism. Nat Cell Biol 2020 22(7):882–895.
    *Corresponding authors: marina.kochetkova@unisa.edu.au, michael.samuel@unisa.edu.au

 Cancer-associated fibroblasts (CAFs) have been             Rho-ROCK pathway with the tumour secretome
reported to exhibit context-dependent tumour-               while revealing a novel tumour-promoting role for
promoting or tumour-suppressing capacities.                 PERK signalling and the enigmatic EGF domain-
However, the mechanisms underpinning these                  containing protein CRELD2.
contexts are not precisely defined. In collaboration          Cancer cells need to modify and remodel the
with other researchers at the Centre for Cancer             environments within which they arise in order to facilitate
Biology, the Walter and Eliza Hall Institute of Medical     their survival, and to enhance their growth and spread.
Research, the Garvan Institute of Medical Research          This is because the normal tissue microenvironment
and the A*STAR Institute of Molecular Cell Biology,         is hostile to the survival and persistence of abnormal
the Samuel lab at the Centre for Cancer Biology             cells. To establish a permissive environment, cancer
has defined a novel signalling axis connecting the          cells therefore need to influence the behaviour of their
VOL 51 NO 3 DECEMBER 2020                     AUSTRALIAN BIOCHEMIST                                            PAGE 13
Publications with Impact

                                                                                             Exposure to Creld2
                                                                                             enhances the
                                                                                             capacity of cancer-
                                                                                             associated fibroblasts
                                                                                             to promote tumour
                                                                                             progression when
                                                                                             they are orthotopically
                                                                                             co-engrafted with
                                                                                             primary mammary
                                                                                             tumour cells into
                                                                                             immune-competent
                                                                                             hosts. Creld2 is
                                                                                             secreted by tumour
                                                                                             cells in which ROCK is
                                                                                             activated, via a novel
                                                                                             mechanism by which
                                                                                             the PERK-regulated arm
                                                                                             of the unfolded protein
                                                                                             response is specifically
                                                                                             engaged in the absence
                                                                                             of ER-stress.

genetically normal microenvironment. Herein lies a key       ECM, the bulk of the tumour ECM is secreted by cells
vulnerability. Since cancer cells need to persistently       populating the microenvironment, such as fibroblasts
maintain their influence over normal cells, interfering      and some immune cells.
with the signals passing between cancer cells and the          To test the hypothesis that cancer cells influence
microenvironment may provide us with opportunities to        the behaviour of fibroblasts via secreted factors, we
normalise the microenvironment such that it is no longer     exposed primary mammary tumour fibroblasts from the
a permissive one.                                            MMTV-PyMT model of mammary cancer to medium
 We had previously demonstrated that the Rho-ROCK            conditioned either by primary mammary tumour cells
signalling pathway, which is frequently activated in         in which the ROCK pathway was activated via the
epithelial cancers, lies upstream of key changes             expression of a construct encoding conditionally active
within the microenvironment. In particular, tumours in       ROCK, or control cells containing a version of the same
which this pathway was activated had high levels of          construct encoding kinase-dead ROCK. Fibroblasts
collagen and other extracellular matrix (ECM) proteins,      exposed to medium conditioned by tumour cells in
enhancing their mechanical stiffness. Elevated tissue        which ROCK had been activated exhibited induction of
stiffness is strongly associated with disease progression,   genes encoding ECM proteins and markers associated
perpetuated by mechano-transduction signalling in            with tumour-promoting CAFs. These fibroblasts were
tumour cells that are located within stiff tissues. These    also more motile than control fibroblasts and cell-
signalling pathways, including signalling via YAP/           derived matrix prepared from them elicited collective
TAZ, beta-catenin and FAK, regulate key processes            and organised migration in tumour cells. Fibroblasts that
that facilitate tumour cell survival, proliferation and      had been exposed to medium conditioned by primary
migration. However, a key question raised by these           mammary tumour cells, in which ROCK was activated,
observations was how Rho-ROCK signalling within              enhanced the growth of tumours when co-engrafted with
tumour cells resulted in the production of large amounts     primary PyMT tumour cells into the mammary fat pads
of ECM proteins. While tumour cells do indeed produce        of wildtype, immune-competent FVB/n female mice.
PAGE 14                                        AUSTRALIAN BIOCHEMIST                       VOL 51 NO 3 DECEMBER 2020
Publications with Impact
 To identify the secreted factors that may be responsible   (ER)-stress. Surprisingly but satisfyingly, recombinant
for inducing the tumour-promoting characteristics in        Creld2 was able to recapitulate the effect of ROCK-
mammary tumour fibroblasts, we conducted proteomic          conditioned medium on mammary tumour fibroblasts,
analysis of the secretome of primary PyMT mammary           rendering them tumour-promoting.
tumour cells in which ROCK had been activated, and            Analysis of signalling downstream of ROCK revealed
compared this with the secretome of primary PyMT            that cells in which ROCK was activated exhibited
mammary tumour cells expressing kinase-dead ROCK.           phosphorylation and activation of Perk, a key sensor
Among the differentially secreted proteins was Creld2,      of ER-stress in one arm of the unfolded protein
a relatively obscure protein that was known to be           response, and indeed that signalling downstream of
secreted by cells subjected to endoplasmic reticulum        Perk was engaged in these cells. Signalling through
                                                            Perk increased the levels and nuclear translocation of
                                                            the transcription factor Atf4, which we demonstrated
                                                            to transcriptionally regulate the gene encoding Creld2.
                                                            This explained the increased production of Creld2 by
                                                            cells in which ROCK was activated.
                                                              Inducing ER-stress in cancers to precipitate UPR-
                                                            mediated immunogenic cell death is an approach that
                                                            is currently the focus of translational research as a
                                                            cancer therapy. Our work therefore serves as a timely
                                                            warning that this approach may yield the unintended
                                                            consequence of enhancing tumour progression via the
                                                            generation of tumour-promoting CAFs.
                                                                             Sarah Boyle, Marina Kochetkova and
                                                                       Michael Samuel, Centre for Cancer Biology
                                                                  SA Pathology and University of South Australia
Sarah Boyle, Marina Kochetkova and Michael Samuel.

         Mutational Landscape of Gall Bladder Cancers
  Pandey A*#, Stawiski EW*#, Durinck S#, Gowda H#, Goldstein LD, Barbhuiya MA, Schroder M,
   Sreenivasamurthy SK, Kim S, Phalke S, Suryamohan K, Lee K, Chakraborty P, Kode V, Shi X,
  Chatterjee A, Datta K, Khan AA, Subbannayya T, Wang J, Chaudhuri S, Gupta S, Srivastav BR,
 Jaiswal BS, Poojary SS, Bhunia S, Garcia P, Bizama C, Rosa L, Kwon W, Kim H, Han Y, Yadav TD,
      Ramprasad VL, Chaudhuri A, Modrusan Z, Roa JC, Tiwari PK, Jang J-Y*, Seshagiri S*.
                 Integrated genomic analysis reveals mutated ELF3 as a potential
                gallbladder cancer vaccine candidate. Nat Commun 2020;11:4225.
                       *Corresponding authors: pandey.akhilesh@mayo.edu,
                   eric.s@medgenome.com, jangjy4@snu.ac.kr, sekar@sgrf.org

 Gall bladder cancer (GBC) is an aggressive                 characterise genomic alterations associated with
gastrointestinal malignancy with a poor prognosis.          GBC. A total of 167 GBC samples and 39 gall bladder
GBC incidence rates are particularly high in specific       non-neoplastic lesions with corresponding matched
geographic regions of the world including Chile,            normal tissue were sequenced. This enabled us to
Bolivia, Peru, Korea, Japan and India. Reasons for          identify genomic alterations frequently observed in
higher incidence rates in these regions remains             GBC and determine if there were differences among
unclear. Women are two to six times more at risk of         tumors from different geographic regions.
GBC than men. The median survival of patients with            We identified several significantly mutated genes
GBC is generally less than a year. This is mainly           that were not previously linked to GBC. This included
because GBC patients are diagnosed at an advanced           ELF3, a frequently mutated gene in GBC with genomic
stage of the disease. Cholelithiasis and cholecystitis      alterations in 21% of tumors. We integrated somatic
are some of the well-known risk factors of GBC.             mutation, copy number variation and gene fusion data to
We established an international collaboration               identify affected pathways in GBC. TP53/RB1 pathway
between researchers from India, USA, South Korea            was most commonly altered in GBC. We also observed
and Chile, and performed whole genome, exome                WNT pathway and KEAP1/NFE2L2 pathway activation
and transcriptome sequencing to identify and                in GBC. Activating mutations in CTNNB1 and RSPO3

VOL 51 NO 3 DECEMBER 2020                     AUSTRALIAN BIOCHEMIST                                        PAGE 15
Publications with Impact
fusion were primary drivers of WNT pathway activation.
We found inactivating mutations in SWI/SNF pathway
genes including SMARCA4, ARID1A and ARID2. We
also identified therapeutically actionable mutations in
RAS/PI3K pathway involving frequent alterations in
ERBB2, ERBB3, BRAF and PIK3CA. About 20% of all
tumors sequenced in our study had actionable mutations
based on approved therapies catalogued in OncoKB.
  Immunotherapy has revolutionised cancer treatment.
Significant survival benefits have been observed
in various cancers including melanoma and lung
cancer. For the first time, we report GBC patients with
mismatch repair defects who have characteristic high
mutation burden and are candidates for check-point
immunotherapy. To further determine the presence of
cancer neoantigens that can be used either alone or in
                                                            Significantly mutated genes in gall bladder cancer.
combination with check-point inhibitors, we evaluated
non-synonymous somatic mutations as candidate               the potential to use cancer immunotherapy for treating
neoantigens. Our analysis identified an average of 15       GBC. Beside serving as a resource for further studying
neoantigens/tumor that were predicted to have high-         GBC and developing therapies, our dataset identified
affinity for MHC class I. Most predicted neoantigens        many actionable GBC alterations that can be treated
were derived from frequently mutated genes such as          using approved therapies.
TP53, ELF3, CTNNB1, ERBB2, ARID1A and CDKN2A.                                                     Harsha Gowda
Using peripheral blood mononuclear cells from HLA-                   QIMR Berghofer Medical Research Institute
matched healthy donors, we determined the ability of
mutant peptides to activate T-cells. Mutant peptides from
ELF3, ERBB2 and TP53 were found to activate T-cells                                                      Left:
suggesting that these peptides are potential cancer                                                      Harsha
vaccine candidates. Some of the ELF3 neoantigens can                                                     Gowda.
be used as a common cancer vaccine candidate as they
are recurrent mutations in GBC and other cancers such                                                    Right:
as colon cancer and ampullary carcinomas.                                                                Somasekar
  Our comprehensive study identifies for the first time                                                  Seshagiri.

                Structural Venomics: from Single to
              Double Knots and Everything in Between
         Pineda SS#*, Chin YKY#, Undheim EAB, Senff S, Mobli M, Dauly C, Escoubas P,
    Nicholson GM, Kass Q, Guo S, Herzig V, Mattick JS, King GF*. Structural venomics reveals
          evolution of a complex venom by duplication and diversification of an ancient
           peptide-encoding gene. Proc Natl Acad Sci USA 2020;117(21):11399–11408.
                                     #Equal first authors
        *Corresponding authors: glenn.king@imb.uq.edu.au, sandy.spineda@gmail.com

 Spiders and other venomous animals depend on               underpinnings of venom complexity. We found that
the production of complex venoms for defence,               the venom of the Australian funnel-web spiders
prey capture and competitor deterrence. Most                evolved primarily by duplication and structural
spider venoms are dominated by disulfide-rich               elaboration a single ancestral knottin gene.
peptides with molecular masses of 3–8 kDa.                   Spiders evolved from an arachnid ancestor around
However, the mechanisms employed by spiders to              450 million years ago (1). Currently, there are more than
generate such chemically and pharmacologically              100,000 extant species of spiders, making them one
diverse venom cocktails is not well understood.             of the most successful animal lineages on the planet.
To address this question, we combined omics and             Like other venomous invertebrates, spiders rely on their
structural biology techniques, a new approach that          venom to capture prey and defend themselves against
we coined ‘structural venomics’, to investigate the         predators. The ongoing battle between venomous

PAGE 16                                       AUSTRALIAN BIOCHEMIST                       VOL 51 NO 3 DECEMBER 2020
Publications with Impact

                                                   Structural innovations in spider venoms knottins. Overview of
                                                   toxin and protein superfamilies highlighting the main mechanisms
                                                   by which toxins have been duplicated, conjugated and
                                                   elaborated upon to form the diversity of knottin scaffolds found
                                                   in venom of the Australian funnel-web spider H. infensa. In all
                                                   structures, disulfide bonds are shown as red tubes, b-sheets and
                                                   a-helices are highlighted in blue and green, respectively.
                                                   Insert: Structural alignment of the core knottin regions of DRPs
                                                   from superfamilies 1 (green), 6 (purple), 17 (pink), 22 (yellow)
                                                   and 23 (orange), highlighting conservation of the knottin motif
                                                   (disulfide-directed b-hairpin in the case of 22) and irrespective of
                                                   the elaborations outside the core region.

animals and their prey and predators places a constant
selection pressure on venom efficacy, and over millions
of years this has resulted in complex arsenals of venom
compounds that vary in size and pharmacological
activity. Spider venoms are dominated by ‘short’ (2–5
kDa) and ‘long’ (6–9 kDa) disulfide-rich peptides
(DRPs). Most spider-venom DRP structures solved to
date correspond to ‘short’ peptides that contain a knottin
motif in which three disulfide bonds are arranged in a
pseudo-knot configuration.
  The venom of the Australian funnel-web spider
Hadronyche infensa is one of the most complex
chemical arsenals in the natural world, comprising
>3000 peptide toxins. We used a combined proteomic,
transcriptomic and structural biology approach to
explore the mechanism by which these spiders evolved
such complex toxin repertoires. We showed that H.
infensa produces 33 superfamilies of venom peptides          From left: Sandy Pineda, Yanni Chin and Glenn King.
and proteins, and that 26 these superfamilies are
DRPs. Moreover, 15 of these DRPs contribute to >90%
of the venom proteome. Structure elucidation using                                                    Sandy Pineda
NMR spectroscopy revealed that most of the DRPs                                  Institute for Molecular Bioscience
are structurally related but present a range of structural                                University of Queensland
innovations. The structural diversity ranged from DRPs                                           Current affilitation:
with a simple knottin motif to highly elaborated knottin                                      Brain and Mind Centre
domains and double-knot domains. Remarkably, this                                              University of Sydney
work enabled us to conclude that the incredible diversity    Reference
of DRPs in the venom of Australian funnel-web spiders        1. Lozano-Fernadez J, et al. (2016) Philos Trans R Soc
is largely derived from a single ancestral knottin gene.        Lond B Biol Sci 371:20150133.

VOL 51 NO 3 DECEMBER 2020                      AUSTRALIAN BIOCHEMIST                                           PAGE 17
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                                                 AUSTRALIAN BIOCHEMIST                    | NO
                                                                                      VOL 51 +613 2DECEMBER
                                                                                                    9541 3500
                                                                                                            2020
ASBMB Education Feature
              The ASBMB Education Feature is coordinated by Nirma Samarawickrema
         (nirma.samarawickrema@monash.edu) and Tracey Kuit (tracey_kuit@uow.edu.au).

                     Online Lessons from Our Students –
                     2020 ASBMB Education Symposium
      Tracey Kuit (University of Wollongong) and Amber Willems-Jones
             (University of Melbourne) interview undergraduate
         Biochemistry students Benjamin Urzua and Rhianna Coscia

 At the recent 2020 ASBMB Education Symposium,                  connected and up-to-date was an important adaptation,
a student panel was held with two undergraduate                 as was talking to my friends for encouragement. This
students: Benjamin Urzua, a third year Bachelor of              helped to put everything in perspective and helped me
Science/Bachelor of Arts student at Monash University           to persevere.
and Rhianna Coscia, a second year Bachelor of Medical           Rhianna – Generally, I adapted well to the transition
Science student at the University of South Australia.           online. A strategy I utilised was to stick to my timetable
Rhianna and Ben shared their experiences learning               as if teaching were on campus. This allowed me to keep
biochemistry online in 2020. Responses to questions             up-to-date with lectures and have tutorial answers ready
have been paraphrased and condensed with permission.            to go for the tutorial Zoom session. The early release
                                                                of lecture recordings was really valuable as I could get
                                                                ahead in my study, allowing ample time for content
                                                                revision at the end of semester.

                                                                What helped you keep engaged online?
                                                                Ben – Online Q&A Zoom sessions after a set of lectures
                                       Biochemistry             were very useful to clarify understanding. Having
                                       students Ben             small group online breakout rooms during the tutorial
                                       Urzua (left) and         sessions, then coming back to the main large group to
                                       Rhianna Coscia.          share our answers. Fresh lectures, rather than the use
                                                                of pre-recorded lectures from previous years, were more
                                                                engaging. Online laboratories remained one of the most
What were the challenges or difficulties with the               engaging aspects of online learning.
rapid move to online learning in 2020?                          Rhianna – I appreciated the recording of classes
Ben – My main difficulties were keeping to a schedule,          delivered online. Whilst I joined classes live, recordings
maintaining a sense of reality and having a sense of            allowed access at any time, and assisted revision.
accomplishment. It was challenging not having the               Though, seeing my tutors or lecturers’ faces whilst
chance to interact with other students, though small            they conveyed the information on Zoom really assisted
group breakout rooms online were the most useful tool           in keeping me engaged, as seeing hand gestures and
to overcome this problem of reduced student interaction.        models provided a human element to what was a very
Rhianna – Challenges included ensuring my home was              technologically driven environment.
suitably set up for a home study environment, quiet and
with reliable internet. An unexpected challenge was the         What were the unexpected benefits of online
difficulty of taking a break from online studies. With all      practical classes?
events cancelled, I found myself spending extended              Ben – Unexpectedly, the practical classes were the
periods of time on university work without a break. As          favourite part of my units. They encouraged me to become
online learning continued, I overcame this by taking            a more active learner in the process and to understand the
regular breaks throughout my day. I felt more focussed          protocols, why each reagent was important and to simply
after incorporating this.                                       be more curious. An online quiz prior to the start of the
                                                                lab ensured we understood the different techniques we
How did you adapt to learning online?                           were going ‘to do’. During the lab, we would go through
Ben – I adapted quite well, though it wasn’t until the          each protocol step while we were asked questions. At
last period of the first semester that I felt that what I had   the end of each session, we had a quiz to complete. This
accomplished so far was due to my own merit. Staying            unexpected benefit in deeper learning may have been

VOL 51 NO 3 DECEMBER 2020                        AUSTRALIAN BIOCHEMIST                                            PAGE 19
ASBMB Education Feature
because we had more time to focus on what we were               Zoom sessions/forums to allow the social interaction and
doing as opposed to rushing to finish everything during a       networking of students, separate to direct class work,
traditional laboratory session.                                 may also assist the online university experience.
Rhianna – Moving practical classes online was naturally
very disappointing, because undertaking a science               What are your recommendations for educators?
degree, the practical, hands on skills are very transferable    Ben – Combining an educational format of self-taught/
to the workplace. However, the transition of practical          self-paced online activities with face-to-face or online
classes to the online environment was successful. We            discussions, where students are encouraged to
had a range of online materials and interactive activities      come prepared with the lecture content and have the
which substituted the face-to-face practicals. This             opportunity to go beyond it. Secondly, having activities
included practical videos which allowed us to visualise         in which students depend on the participation of other
what we would have been doing in the practical session,         students, e.g. there could be some group activities where
but also provided greater understanding to how supplied         each student needs to contribute something by having
sample results were obtained from recipe-like steps in a        completed a task prior to the lecture/tutorial. This could
lab book.                                                       help students to become more active learners and to
                                                                collaborate.
What were your frustrations with online learning?               Rhianna – Continuing to offer online interactive activities
Ben – I understand that collusion can be an issue and           to supplement face-to-face learning, as it provides
that having your notes by your side is an impediment            instantaneous feedback about content understanding,
to reliably measure how students are able to apply the          and allows access at time of need. Secondly, consider
unit’s content. However, sometimes test time constraints        communication between teaching staff and students a
meant we didn’t have enough time to show our true               key importance when delivering learning online. There
potential. Additionally, it was frustrating not being able to   have been occasions in some classes where we have
draw reaction mechanisms or biochemical pathways.               not been contacted by educators, leaving students in
Rhianna – Rather than a frustration as such, the                limbo about what to do, what time to log on etc., causing
reliability of the online assessment platform was anxiety       undue stress. Thirdly, adhere to the timetable, utilising
inducing. For example, in one particular test, my internet      visual elements and interactive activities (e.g. Kahoot) to
connection dropped out due to the number of students            maintain the focus of students who are at home.
logging on simultaneously. This caused additional anxiety
in a test setting. There are technological aspects of online
learning (i.e. countdown time limits, file compatibility,
etc.) that can occasionally be unreliable.                          Tracey Kuit is an Education-
                                                                   Focused Associate Professor
What things would you change?                                         in the School of Chemistry
Ben – Having more instances for students to engage                    and Molecular Bioscience,
with one another. For example, providing online readings               University of Wollongong.
or short videos related to lectures and giving students                 tracey_kuit@uow.edu.au
a chance to discuss with the lecturer and develop their
critical thinking skills. I would be cautious, however, not
to expect too much from students, because sometimes
it felt that because students were at home, academics
assumed they had more time and therefore gave an                      Dr Amber Willems-Jones
increased workload as a result.                                      is a Senior Lecturer in the
Rhianna – As someone that likes to be organised and get            Department of Biochemistry
ahead in my studies, I value the upload of content early                 and Molecular Biology,
to have efficient revision time at the end of semester.                University of Melbourne.
Online discussion forums also permit students to interact       amber.willems@unimelb.edu.au
with one another about content-related questions. Some

PAGE 20                                          AUSTRALIAN BIOCHEMIST                         VOL 51 NO 3 DECEMBER 2020
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