Newsletter Department of Biochemistry and Molecular Biology

www.med.monash.edu.au/biochem

Issue 15: August 2011

NEWS AND EVENTS

Nature Immunology Advance Online PublicationSeminars in September

4 pm on Wednesdays in Building 13A, Lecture Theatre M2

September 7 Edwina McGlinn (ARMI)

The Role of miR-196 in Vertebrate Development

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September 14 Michael Cowley (Physiology Dept)

TBA

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September 21 Lisa Martin (Monash, Chemistry Dept)

Salt, Sugar & Sex

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September 28 Mark Wright (Monash, Immunology)

The Tetraspanin CD37 is Essential for Integrin Function and Immune Cell Migration

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Please send suggestions for future speakers

to committee members: Travis Beddoe, Melanie Pritchard,

Martin Stone, Ana Traven, Catherine Itman, Peter Boag

A list of all seminars for Semester 2 can be found on the Biochem webpage

www.med.monash.edu.au/biochem under “About”

Recognition of β-linked self glycolipids mediated by natural killer T cell antigen receptors

Daniel G Pellicci, Andrew J Clarke, Onisha Patel, Thierry Mallevaey, Travis Beddoe, Jérôme Le Nours, Adam P Uldrich, James McCluskey, Gurdyal S Besra, Steven A Porcelli, Laurent Gapin, Dale I Godfrey & Jamie Rossjohn

Abstract:

The most potent foreign antigens for natural killer T cells (NKT cells) are α-linked glycolipids, whereas NKT cell self-reactivity involves weaker recognition of structurally distinct β-linked glycolipid antigens. Here we provide the mechanism for the autoreactivity of T cell antigen receptors (TCRs) on NKT cells to the mono- and tri-glycosylated β-linked agonists β-galactosylceramide (β-GalCer) and isoglobotrihexosylceramide (iGb3), respectively. In binding these disparate antigens, the NKT cell TCRs docked onto CD1d similarly, achieving this by flattening the conformation of the β-linked ligands regardless of the size of the glycosyl head group. Unexpectedly, the antigenicity of iGb3 was attributable to its terminal sugar group making compensatory interactions with CD1d. Thus, the NKT cell TCR molds the β-linked self ligands to resemble the conformation of foreign α-linked ligands, which shows that induced-fit molecular mimicry can underpin the self-reactivity of NKT cell TCRs to β-linked antigens.

LATEST MEMBERS OF STAFF

Lab Head New Staff Member Position

Prof Trevor Lithgow Hsin-Hui Shen ARC Super Science

Prof David Jans Johanna Dean Research Fellow

Prof David Jans Kim Lieu Research Assistant

Prof Tony Tiganis Esteban Gurzov Research Fellow

Page 2Newsletter: August 2011, Issue 15 Department of Biochemistry and Molecular Biology

ATTENTION ALL PEOPLE HANDLING/USING ANIMALS AT MONASH UNIVERSITY

Staff and Students are required to attend an Information Session and a Practical Training session on the use and handling of animals. This is compulsory for those who will be involved in animal research. All users must be familiar with the current regulations regarding animal care and use in research and teaching within the university. If unsure which training session would best suit you, contact your Deptl Safety Officer (Irene Hatzinisiriou) or BioSafety Officer for advise. There are specific guidelines to adhere to when working with animals, and also adequate training records must be kept within the department for all future Safety/Bio-Safety Audits.

All animal injuries to humans no matter how trivial they may seem, should be reported to a First Aider and Safety Officer as soon as they occur. The necessary medical advise/intervention will be given by trained medical staff for any such incidents. If an animal injury is left untreated, it may become a more serious medical issue over time.

Protect yourself and report it immediately!

WEARING THE CORRECT PERSONAL PROTECTION EQUIPMENT (PPE) IN THE LAB!

If you are unsure as to what type of PPE you should be wearing in the lab, here are a few steps to work it out!

1. Always wear your lab coat when carrying out any procedure in the lab

2. Always wear the correct footwear (closed shoes, no bare skin showing)

3. Read through your protocol/procedure carefully

4. Read the associated MSDS

5. Read the associated risk assessment, as this will guide you to as to WHAT type of GLOVES and/or SAFETY EYE WEAR you should be wearing when carrying out this protocol/procedure

6. Risk assessment will also indicate WHERE you should be carrying out this protocol/procedure

WEARING THE CORRECT GLOVES!

There are gloves specific for most processes which are carried out in our labs.

Please use the correct gloves (and correct size), as listed in the risk assessments.

All gloves should be disposed of in the Biohazard or Cytotoxic bins. Do not place any gloves in any landfill bin!

Latex gloves

• offer a temporary barrier between you and your samples

• should be regularly changed

• when soiled should be removed asap

• avoid touching doors, telephones and keyboards with gloves on

Nitrile gloves

• offer a barrier to acids and aliphatic hydrocarbons

• are less flexible than latex gloves

• are more puncture-resistant than latex gloves

• should not be worn when handling aromatic hydrocarbons, esters, aldehydes and ketones

• should be regularly changed

• when soiled should be removed ASAP

• avoid touching doors, telephones and keyboards with gloves on

Cotton gloves

• used for keeping your fingers warm

• wear a pair of latex gloves over them to keep them dry when handling cold samples

Cryogenic “SPECIAL” gloves (not leather, rubber or gardening gloves)

• used for handling/storing samples in and out of Liquid Nitrogen

• used in conjunction with a face shield!

Page 3Department of Biochemistry and Molecular Biology

SPOTLIGHT ON: Allyson Croxford

After completing a Bachelor of Science degree at Monash University, majoring in Biochemistry and Marine Biology, I went on to complete a summer scholarship followed by an Honours year in the Department of Biochemistry and Molecular Biology under the supervision of A/Prof Merrill Rowley. After a successful honours year I went on to study a PhD under the co-supervision of A/Prof. Merrill Rowley from the Department of Biochemistry and Molecular Biology and Prof. Don McNaughton from the School of Chemistry.

My PhD focuses on the development of infrared microspectroscopy techniques to examine the effect of monoclonal antibodies to type II collagen in a murine model of rheumatoid arthritis. The aim of my research was to investigate the effect of combinations of known arthritogenic antibodies to type II collagen on pre-existing cartilage both in vitro on bovine cartilage explants and in vivo in mice that lack inflammation. I have been lucky enough to undertake part of my research on the infrared beamline at the Australian Synchrotron. The use of standard histological techniques, laboratory infrared spectroscopy and synchrotron infrared microspectroscopy has provided evidence for the direct destructive effects of antibodies to cartilage collagen on pre-existing cartilage, in the absence of inflammation. A striking observation which has resulted from my studies is the demonstration that not only can antibodies to type II collagen have destructive effects on pre-existing cartilage both in vitro and in vivo but one antibody that does not have any effect alone, is protective

against the degradative effects caused by the other antibodies. Since antibodies of the same specificity to those used in my studies have been associated with human arthritis, my studies have important implications in the overall understanding of the mechanisms which occur in human rheumatoid arthritis. Furthermore, understanding the protective mechanism of the protective antibody may be important in the development of new treatments for RA. The use of infrared spectroscopy, particularly at the Australian synchrotron has identified changes to the cells and cartilage matrix before the onset of macroscopic inflammation. The applicability of infrared spectroscopy for analysis of cartilage should also be useful in other cartilage diseases such as osteoarthritis, in cartilage replacement therapies and development of biomaterials.

I have been lucky enough to have a successful PhD. In 2009 I won the inaugural Monash University 3 minute thesis competition for outlining my research. This was a huge boost for me as I originally entered just to have a go at summarising my PhD in less than 3 minutes and never thought I would take out the overall prize from Monash University. It was such an honour to win for studying rheumatoid arthritis, a disease which is close to my heart, as I have it myself. During the course of my PhD, I have also won prizes at conferences for my posters and I have successfully published two papers investigating the effect of combinations of mAbs to type II collagen on pre-existing cartilage both in vitro and in vivo. I have just submitted my PhD thesis for examination.

The 2011 Biochemistry Postgraduate Research Conference will be held on the 10th and 11th of November.

The Conference provides a platform for all PhD students to present their work to the department. As such, all postgraduate students are invited and highly encouraged to present posters at the conference.

Abstracts for posters should be maximum 300 words, and the posters themselves no larger than 90cm x 120cm (Velcro attachments will be provided on the day!). There will be great prizes for best posters, so don’t miss this opportunity and start working on your abstracts now!

Please submit abstracts for your poster by the 16th of September to Caitlin Rowe (caitlin.rowe@monash.edu) or Linda Wiltzer (linda.wiltzer@monash.edu).

The conference is always a lot of fun, so join in, and enter the 10th and 11th of November in your diaries as days that cannot possibly be missed!

2011 Biochemistry Postgraduate Research Conference

Environmental Sustainability At MonashAnyone concerned with any environmental issues should either contact Dalibor.Mijaljica@monash.edu or visit The Office of Environmental Sustainability (TOES) http://www.fsd.monash.edu.au/environmental-sustainability.

Everyone is welcome to take the Monash Environmental Pledge 2011 http://fsd.monash.edu.au/environmental-sustainability/monash-environmental-pledge-2011

Newsletter: August 2011, Issue 15

Page 4Department of Biochemistry and Molecular BiologyNewsletter: August 2011, Issue15

RECENT PUBLICATIONS

1Ambaye, N.D., Pero, S., Gunzburg, M.J., Yap, M.Y., Clayton, D.J., Del Borgo, M.P., Perlmutter, P., Aguilar, M.I., Shukla, G.S., Peletskaya, E., Cookson, M.M., Krag, D.N., Wilce, M.C., and Wilce, J.A., Structural basis of binding by cyclic non-phosphorylated peptide antagonists of GRB7 implicated in breast cancer progression. J Mol Biol 2011. E-pub: p. 1-42

2 Archbold, J.K., Flanagan, J.U., Watkins, H.A., Gingell, J.J., and Hay, D.L., REVIEW:Structural insights into RAMP modification of secretin family G protein-coupled receptors: implications for drug development. Trends Pharmacol Sci 2011. E-pub: p. 1-10

3 Chow, J.D., Jones, M.E., Prelle, K., Simpson, E., and Boon, W.C., A Selective Estrogen Receptor Alpha Agonist Ameliorates Hepatic Steatosis in the Male Aromatase Knockout Mouse. J Endocrinol 2011. E-pub: p. 1-39

4 D’Cruze, T., Gong, L., Treerat, P., Ramm, G., Boyce, J., Prescott, M., Adler, B., and Devenish, R.J., A role for the Burkholderia pseudomallei Type Three Secretion System cluster 1 bpscN gene in virulence. Infect Immun 2011. E-pub: p. 1-26

5Iglesias, M.C., Almeida, J.R., Fastenackels, S., van Bockel, D.J., Hashimoto, M., Venturi, V., Gostick, E., Urrutia, A., Wooldridge, L., Clement, M., Gras, S., Wilmann, P.G., Autran, B., Moris, A., Rossjohn, J., Davenport, M.P., Takiguchi, M., Brander, C., Douek, D.C., Kelleher, A.D., Price, D.A., and Appay, V., Escape from highly effective public CD8+ T-cell clonotypes by HIV. Blood 2011. E-pub: p. 1-38

6 Lapinskas, E.J., Svobodova, S., Davis, I.D., Cebon, J., Hertzog, P.J., and Pritchard, M.A., The Ets Transcription Factor ELF5 Functions as a Tumor Suppressor in the Kidney. Twin Res Hum Genet 2011. 14(4): p. 316-22

7 Mijaljica, D., Prescott, M., and Devenish, R.J., VIDEO: A fluorescence microscopy assay for monitoring mitophagy in the yeast Saccharomyces cerevisiae. J Vis Sci 2011. On-Line: p. 1-5

8 Lagiakos, H.R., Walker, A., Aguilar, M.I., and Perlmutter, P., Thionation of amides using a solid-supported P2S5 reagent under microwave irradiation. Tetrahedron Lett 2011. E-pub: p. 1-1-5

9 Phan, J., Yamout, N., Schmidberger, J., Bottomley, S.P., and Buckle, A.M., Refolding Your Protein with a Little Help from REFOLD, in Methods Mol Biol, KJ Barnham AF Hill, SP Bottomley & R Cappai, Editor. 2011, Humana Press: UK. p. 45-57. Book Chapter [BCH]

10 Saunders, H.M., Gilis, D., Rooman, M., Dehouck, Y., Robertson, A.L., and Bottomley, S.P., Flanking domain stability modulates the aggregation kinetics of a polyglutamine disease protein. Protein Sci 2011. E-pub: p. 1-23

11 Zhai, J., Wooster, T.J., Hoffmann, S.V., Lee, T.H., Augustin, M.A., and 2011, Structural Rearrangement of beta-Lactoglobulin at Different Oil-Water Interfaces and Its Effect on Emulsion Stability. Langmuir 2011. E-pub: p. 1-10

POSTGRADUATE MATTERS

Information SessionProf Mibel Aguilar will be presenting a information session on applying for a scholarship to the Hons students on Fri Sept 2 at 11.30 in Rm 204/205. Anyone else interested is welcome to attend.

Scholarship ApplicationsThe next Round of scholarship applications close October 31. http://www.mrgs.monash.edu.au/scholarships/apply/

Link to MBio e-bulletin: http://www.med.monash.edu.au/mbio-gradschool/e-bulletin.html

To find out more about NOTDRS please visit: http://www.med.monash.edu.au/biochem/student-society.html

Or find us on facebook: http://www.facebook.com/groups.php?id=695417331&gv=12#/group.php?gid=86877543416

All queries on Postgraduate matters: Please contact Prof Mibel Aguilar