Peptide Laboratory
Laboratory Head
A/Prof John D Wade BSc (Hons) PhD FRACI FRSC
Contact Details
Email: | |
Phone: | +61 (0)3 8344 7285 |
Fax: | +61 (0)3 9348 1707 |
Number of
Staff: 5
- Dr Eleni Giannakis eleni.giannakis@florey.edu.au
- Dr Akhter Hossain akhter.hossain@florey.edu.au
- Dr Suode Zhang suode.zhang@florey.edu.au
- Mr Feng Lin feng.lin@florey.edu.au
- Mrs Mary Macris mary.macris@florey.edu.au
Students: 2
- Lin-Wai Hung
- Fazel Shabanpoor
Research Interests
The principal focus of our laboratory is the structure and function relationship of members of the insulin superfamily, particularly relaxins-2 and 3 and insulin-like peptide 3 (INSL3). We are endeavouring to understand how their three dimensional shape dictates their unique biological properties and to use this information to design and construct mimetics of these peptides that may have therapeutic applications.
Relaxin and INSL3 each consists of two peptide chains, A and B, that are held together by three disulfide bonds, one within the A-chain and two intermolecular between the two chains. In humans, there are two relaxins known as 2 and 3. Relaxin-2 is produced principally in the ovaries of preganant females and has important post-partum roles. Relaxin-3 was recently discovered at the Florey and appears to be a neuropeptide having key roles in stress and appetite regulation. INSL3 has a primary role in initiating testes descent in the developing male and in the adult is an important component of the gonadotropic axis and regulates germ cell maturation.
We use state-of-the-art peptide synthesis technology to prepare these peptide hormones in sufficient quantities to allow their detailed biochemical study, that is, how they bind to their receptors to cause their unique biological effects. We also use drug design techniques in conjunction with tertiary structural techniques to prepare analogues of the peptides that are predicted to be more potent, longer lasting and specific.
We also undertake detailed protein profiling and biomarker discovery with the goal of using this technology to study the role of these peptides in health and disease. Towards this aim, use a Ciphergen surface-enhanced laser desorption/ionization (SELDI) ProteinChip System. It is a multi-dimensional protein recognition tool and structure analyser. It enables protein capture, purification, analysis, and processing from complex biological mixtures directly on derivatised surfaces. Peptides and proteins are affinity-immobilised onto the surface of chemically or biochemically activated ProteinChips. Retained analytes are detected according to molecular weight and quantitated by the use of a time-of-flight mass spectrometer (TOF MS) equipped with a laser desorption ion source. We have also been using this technology to understand the role of amyloid-beta in neurological disease and have logically extended the work to include other areas of neuroscience.
Current Projects
- Determining the structure and function relationships of human relaxin-2 and -3 and insulin-like peptide 3 (Akhter Hossain, Feng Lin, Fazel Shabanpoor, Suode Zhang, John Wade with Tony Hughes, Department of Pharmacology, University of Melbourne).
- Protein profiling of relaxin KO mice, a model of progressive fibrosis (Eleni Giannakis, Chrishan Samuel, Mary Macris, Geoffrey Tregear, John Wade).
- Capture and quantitation of Abeta, a marker for Alzheimer's Disease (Eleni Giannakis, John Wade and Lin Hung with Kevin Barnham, Keyla Perez and Colin Masters, Department of Pathology, University of Melbourne).
- The mechanism of binding of Abeta to cell membranes (Keyla Perez with Frances Separovic, Bio21 Institute, University of Melbourne).
- Capture and quantitation of alpha-synuclein, a marker for Parkinson’s Disease (Eleni Giannakis and John Wade with Rogan Tinsley, Janelle Nunan and Mal Horne).
- Protein profiling of D2 KO mice, a model of Parkinson’s Disease (Eleni Giannakis, Mary Macris and John Wade with Janelle Nunan, Rogan Tinsley and Mal Horne).
- Design and development of novel antibacterial peptides (John Wade and Feng Lin with Laszlo Otvos, Temple University, Philadelphia, USA, and Ralf Hoffman, Leipzig University, Germany).
- Mining reptile venom for new peptide therapeutics (John Wade and Feng Lin with Bryan Greig Fry, Australian Venom Research Unit, University of Melbourne).
- The structure and function of insulin analogues as potential new diabetes treatments (Akhter Hossain, Feng Lin, John Wade with Andrea Robinson, School of Chemistry, Monash University).
Laboratory Techniques
- Solid phase peptide synthesis
- Reversed phase-HPLC
- MALDI-TOF and SELDI-TOF mass spectrometry
- Computer-aided drug design
- Amino acid analysis
- Peptide engineering
Funding
- National Health and Medical Research Council (Australia) - Project grants
- Australian Research Council Discovery grant
- Diabetes Australia
- The Ian Potter Foundation
- The R.E. Ross Foundation
Additional Information
Recent Peptide Laboratory publications
Agbottah, E., Zhang, N., Dadgar, S., Pumfery, A., Wade, J.D., Zeng, C. and Kashanchi, F. (2006) Inhibition of HIV-1 virus replication using small soluble Tat peptides. Virology, 345, 373-389.
Bathgate, R.A.D., Lin, F., Hanson, N.F., Otvos, Jr. L., Guidolin, A., Giannakis, C., Bastiras, S., Layfield, S., Ferraro, T., Ma, S., Zhao, C., Gundlach, A.L., Samuel, C.S., Tregear, G.W. and Wade, J.D. (2006) Relaxin-3: Improved synthesis strategy and demonstration of its high-affinity interaction with the relaxin receptor LGR7 both in vitro and in vivo. Biochemistry, 45, 1043-1053.
Rosengren, K.J., Lin, F., Bathgate, R.A.D., Wade, J.D. and Craik, D.J. (2006) Solution structure and insights into the determinants of receptor specificity of human relaxin-3. Journal of Biological Chemistry, 281, 5845-5851.
Berro R, Kehn K, de la Fuente C, Pumfery A, Adair R, Wade JD, Colberg-Poley AM, Hiscott J and Kashanchi F. (2006) Acetylated TAT regulates HIV-1 splicing through its interaction with the splicing regulator, p32. Journal of Virology 80, 3189-3204.
Wilson, L.M., Pham, C.L.L., Jenkins, A.J., Wade, J.D., Hill, A.F., Perugini, M.A. and Howlett, G.J. (2006) High density lipoproteins bind amyloid- and apolipoprotein C-II amyloid fibrils. Journal of Lipid Research, 47, 755-760.
Fu, P., Shen, P-J., Zhao, C-X., Scott, D.J., Samuel, C.S., Wade, J.D., Tregear, G.W., Bathgate, R.A. and Gundlach, A.L. LGR8 (RXFP2) in mature glomeruli of developing and adult rat kidney and inhibition by INSL3 of glomerular cell proliferation. Journal of Endocrinology, 189, 397-408.
Del Borgo, M., Hughes, A.R., Lin, F., Bathgate, R.A.D., Kawamura, K. and Wade, J.D. Conformationally constrained mimetics of insulin-like peptide 3 B-chain are specific LGR8 antagonists. Journal of Biological Chemistry, 281, 13068-13074, 2006.
Hossain, A., Lin, F., Zhang, S., Ferraro, T., Bathgate, R.A.D., Tregear, G.W. and Wade, J.D. Regioselective disulfide solid phase synthesis, chemical characterization and in vitro receptor binding activity of equine relaxin. International Journal of Peptide Research & Therapeutics, 12, 211-215, 2006.
Smith, D.P., Smith, D.G., Curtain, C.C., Boas, J.F., Pilbrow, J.R., Ciccotosto, G.D., Lau, T-L., Tew, D.J., Perez, K., Wade, J.D., Bush, A.I., Drew, S.C., Separovic, F., Masters, C.L., Cappai, R. and Barnham, K.J. Copper-mediated amyloid-beta toxicity is associated with an intermolecular histidine bridge. Journal of Biological Chemistry, 281, 15145-15154, 2006.
Otvos, Jr., L., De Olivier Inacio, O., Wade, J.D. and Cudic, P. (2006) Prior antibacterial peptide-mediated inhibition of protein folding in bacteria mutes resistances enzymes. Antimicrobial Agents and Chemotherapy, 50, 3146-3149, 2006.
Rosengren, K.J., Zhang, S., Lin, F., Daly, N.L., Scott, D.J., Hughes, R.A., Bathgate, R.A.D., Craik, D.J. and Wade, J.D. Solution structure and characterization of the receptor binding surface of insulin-like peptide 3. Journal of Biological Chemistry, 281, 28287-28295, 2006.
Shabanpoor, F., Bathgate, R.A.D., Hossain, M.A., Giannakis, E., Wade, J.D. and Hughes, R.A. Design, synthesis and evaluation of cyclic analogues of insulin-like peptide 3. Journal of Peptide Science (accepted for publication 1 October 2006).
Research
