Skip to the content
 

Associate Professor Martin Stone

Martin Stone

Telephone:
+61 - 3 9905 9537

Facsimile:
+61 - 3 9905 3726

Email:
Martin.Stone@med.monash.edu.au

Brief Biosketch

  • B.Sc., University of Auckland, New Zealand (1986)
  • M.Sc.(hons), University of Auckland, New Zealand (1987)
  • Ph.D., University of Cambridge, UK (1991)
  • Research Associate, The Scripps Research Institute, USA (1991-1995)
  • Assistant Professor, Indiana University, USA (1995-2002)
  • Associate Professor, Indiana University, USA (2002-2007)
  • Associate Professor, Monash University, Australia (2007-)

Chemokine Receptors in Inflammation and Infection

Inflammation is the response of a tissue and its microvascular system to injury or infection. A hallmark of inflammation is the accumulation of leukocytes (white blood cells), which remove pathogens and necrotic tissue. However, excessive leukocyte recruitment can lead to degradation of healthy tissue, i.e. inflammatory disease. Leukocyte recruitment in inflammation is controlled by the expression and secretion of small proteins called chemokines at the site of inflammation and by the subsequent interaction of those chemokines with chemokine receptors located on the surfaces of circulating leucocytes. A detailed understanding of chemokine-receptor interactions is required in order to rationally develop novel therapeutic agents against inflammatory diseases.

In addition to their roles in inflammation, chemokine receptors play critical functions in infection of blood cells by pathogens. In particular, the AIDS virus HIV-1 binds uto chemokine receptors in order to gain entry into leukocytes and the malarial parasite Plasmodium vivax hijacks a chemokine receptor to invade red blood cells. Detailed molecular studies of these interactions will assist efforts to prevent and cure these major infectious diseases.

Project areas

Structural Basis of Chemokine-Receptor Recognition: a Protein Engineering Approach

The intricate coordination of leukocyte trafficking (migration to the right tissues at the right times) is controlled, to a large extent, by the specific interactions between the chemokines and their receptors. For this reason, we have a major research effort towards understanding the structural basis of these specific interactions. This is particularly challenging because chemokine receptors are G protein-coupled receptors (integral membrane proteins) making them difficult to isolate or characterize in detail. Our studies utilize a wide variety of approaches including: NMR studies of chemokine structure; binding and activity studies of mutant and chimeric chemokines and receptors; and peptide models of the receptors.

   Receptor peptide binding site highlighted on the structure of the chemokine eotaxin

Receptor peptide binding site highlighted on the structure of the chemokine eotaxin

Second diagram

We have developed a family of novel proteins (dubbed "CROSS" proteins) in which chemokine-binding elements of the receptors are displayed on the surface of a soluble protein scaffold. The CROSS proteins bind to chemokines with the correct specificity and therefore can be used to determine the detailed structural interactions giving rise to specific recognition. A major goal of our current research is to determine the structures of the chemokine-CROSS complexes.

Malarial Infection via the Duffy Antigen Receptor for Chemokines (DARC)

Malaria is a mosquito-transmitted parasitic disease that affects more than half a billion people worldwide each year and has a devastating effect on both public health and economic prosperity in numerous countries, especially those in equatorial regions. The pathogenic organisms in malaria are several species from the protozoan genus Plasmodium. One of these species, Plasmodium vivax, infects human reticulocytes (immature red blood cells) by docking with the chemokine receptor DARC on the reticulocyte surface. Building on our soluble mimics of chemokine receptors, we are engineering soluble proteins derived from DARC in order to characterize the interactions between DARC and the Plasmodium vivax Duffy-binding protein (DBP). These studies will identify essential elements of DBP, thereby guiding the development of vaccines to prevent malarial infection.

The Role of Tyrosine Sulfation in Chemokine Receptor Recognition by Chemokines and Malarial Parasites

The interactions of chemokine receptors with both chemokines and Duffy binding protein are enhanced by post-translational sulfation of critical tyrosine residues located in the extracellular amino-terminal regions of the receptors. However, detailed molecular studies of the influence of sulfation have been limited to date by the integral membrane nature of the receptors as well as the difficulty obtaining homogeneously sulfated proteins. Using a combination of synthetic and enzymatic approaches, we are now investigating the role of sulfation in controlling the binding of chemokine receptors to chemokines and DBP. In addition to enhancing our understanding of these specific systems, our studies will provide fundamental insights into how Tyr sulfation can be used to regulate protein-protein recognition.

   Diagram of sulfotyrosine

Sulfotyrosine

Potential tyrosine sulfation sites in some chemokine receptors

Potential tyrosine sulfation sites in some chemokine receptors

Research and Training Opportunities

We are enthusiastic to recruit Honours and Ph.D. students with interests in biochemistry, biomedical science and chemistry to participate in our ongoing research projects.  Research students receive training in a wide variety of skill areas including:

  • Molecular biology (cloning, mutagenesis, sequencing, etc.)
  • Protein expression and purification
  • Peptide synthesis
  • Analytical biochemistry methods
  • Binding measurements
  • Physical and spectroscopic techniques (fluorescence, calorimetry, circular dichroism, etc.)
  • Protein structural analysis by NMR and X-ray crystallography
  • Data analysis methods
  • Bioinformatics and computational methods

Research Funding

Past and present research in the laboratory has been funded from the following sources:

  • National Institutes of Health (USA)
  • National Science Foundation (USA)
  • American Heart Association
  • American Chemical Society
  • Australian Research Council

Selected Publications

  • Jarymowycz, V.A., Cortajarena, A.L., Regan, L., and Stone, M.J. Comparison of the backbone dynamics of a natural and a consensus designed 3-TPR domain. J. Biomol. NMR (2008) 41, 169-178.
  • Krop, E.J.M., Matsui, E.C., Sharrow, S.D., Stone, M.J., Gerber, P., and Aalberse, R.C.  Recombinant major urinary proteins of the mouse in specific IgE and IgG testing. International Archives of Allergy and Immunology (2007) 144, 296-304.
  • Krop, E.J.M., Doekes, G., Stone, M.J., Aalberse, R.C., and van der Zee, J.S.  Spreading of occupational allergens: laboratory animal allergens on hair covering caps and in mattress dust of laboratory animal workers.  Occupational and Environmental Medicine (2007) 64, 267-272.
  • Cheng, C.Y., Jarymowycz, V.A., Cortajarena, A.L., Regan, L., and Stone, M.J.  Domain Motions and Backbone Flexibility in Designed Proteins with Different Numbers of Identical Consensus Tetratricopeptide Repeats.  Biochemistry (2006) 45, 12173-12183.
  • Jarymowycz, V.A., Krupinska, E., and Stone, M.J.  Comparison Between the Backbone Dynamics of an 11-Amino Acid Peptide Sequence in Alpha-Helical and Beta-Hairpin Structural Contexts. Biochemistry (2006) 45, 11179-11189.
  • Jarymowycz, V.A. and Stone, M.J.  Fast Time Scale Dynamics of Protein Backbones: NMR Relaxation Methods, Applications, and Functional Consequences.  Chemical Reviews (2006) 106, 1624-1671.
  • Sharrow, S.D., Edmonds, K.A., Goodman, M.A., Novotny, M.V., and Stone, M.J.  Thermodynamic Consequences of Disrupting a Water-Mediated Hydrogen Bond Network in a Protein:Pheromone Complex.  Protein Science (2005) 14, 249–256.
  • Goehlert, V.A., Krupinska, E., Regan, L., and Stone, M.J.  Analysis of Side Chain Mobility among Protein G B1 Domain Mutants with Widely Varying Stabilities.  Protein Science (2004) 13, 3322–3330.
  • Datta-Mannan, A. and Stone, M.J.  Chemokine Binding Specificity of Soluble Chemokine Receptor Analogues: Identification of Interacting Elements by Chimera Complementation. Biochemistry (2004) 43, 14602-14611
  • Mayer, M.R., Parody, T.R., Datta-Mannan, A., and Stone, M.J.  Specificity Determinants for Chemokine Recognition Identified Using Eotaxin-MCP-1 Chimeras. FEBS Lett. (2004) 571, 166-170.
  • Parody, T.R. and Stone, M.J.  Activation, and Antagonism of CC Chemokine Receptors CCR2 and CCR3 in Chinese Hamster Ovary Cells.  Cytokine (2004) 27, 38-48.
  • Datta, A. and Stone, M.J.  Soluble Mimics of a Chemokine Receptor: Chemokine Binding by Receptor Elements Juxtaposed on a Soluble Scaffold.  Protein Science (2003) 12, 2482-2491.
  • Mayer, K.L., Earley, M.R., Gupta, S., Pichumani, K., Regan, L., and Stone, M.J.  Covariation of Backbone Motion Throughout a Small Protein Domain.  Nature Structural Biology (2003) 10, 962-965
  • Sharrow, S.D., Novotny, M.V., and Stone, M.J. Thermodynamic Analysis of Binding Between Mouse Major Urinary Protein-I and the Pheromone of 2-sec-Butyl-dihydrothiazole. Biochemistry (2003) 42, 6302-6309
  • Mayer, K.L. and Stone, M.J. Backbone Dynamics of the CC-Chemokine Eotaxin-2 and Comparison Among the Eotaxin Group Chemokines. Proteins: Structure, Function and Genetics (2003) 50, 184-191.
  • Sharrow, S.D., Vaughn, J.L., Zidek, L., Novotny, M.V., and Stone, M.J.  Pheromone Binding by Polymorphic Mouse Major Urinary Proteins.  Protein Science (2002) 11, 2247-2256.
  • Ye, J., Mayer, K.L., Mayer, M. R., and Stone, M.J.  NMR Solution Structure and Backbone Dynamics of the CC-Chemokine Eotaxin-3.  Biochemistry (2001) 40, 7820-7831.
  • Mayer, M.R. and Stone, M.J.  Identification of Receptor Binding and Activation Determinants in the N-Terminal and N-Loop Regions of the CC Chemokine Eotaxin.  J. Biol. Chem. (2001) 276, 13911-13916.
  • Ye, J., Kohli, L.L., and Stone, M.J.  Characterization of Binding Between the Chemokine Eotaxin and Peptides Derived from the Chemokine Receptor CCR3.  J. Biol. Chem. (2000) 275, 27250-27257.
  • Mayer, K.L. and Stone, M.J.  NMR Solution Structure and Receptor Peptide Binding of the CC Chemokine Eotaxin-2.  Biochemistry (2000) 39, 8382-8395.
  • Zídek, L., Novotny, M.V., & Stone, M.J.   Increased protein backbone conformational entropy upon hydrophobic ligand binding.  Nature Structural Biology (1999) 6, 1118-1121.
  
Staff Listings