Structural and Functional Proteomics of Bacterial Pathogens

Main research interests:

Biology of pathogenic bacteria:
Our major interest is focused on understanding the strategies employed by intracellular pathogens like Salmonella, Legionella or Chlamydia for infection and persistence within human hosts. Our integrated research platform utilizes a combination of approaches including proteomics-based target identification, molecular characterization of potentially critical host-pathogen interactions, structural elucidation of identified targets and macromolecular complexes by X-ray crystallography, and structure-based drug design of novel antimicrobials

Central technologies:

Proteome analysis of intracellular pathogens:
The eukaryotic and prokaryotic proteins necessary for intracellular bacterial survival are analysed by 2DE and two dimensional chromatography. Comparison of different infection models, host variation and and mutant bacteria will allow another insight into their intracellular life style.
Protein characterization by mass spectrometry:
Along with the structural and functional analysis of virulence-associated proteins, Nano-LC ESI mass spectrometry will be used for the analysis of complex mixtures (such as peptide digests), assessment of protein purity, protein sequencing and detection of post-translational modifications (mass spectrometry facility planned for 2006).
Structural analysis of bacterial drug targets:
X-ray crystallography is an indispensable tool for illuminating the structural principles underlying biological phenomena such as intracellular infection. We've implemented a structural proteomics strategy involving focused X-ray crystallographic studies on promising targets and macromolecular complexes identified by complementary methods. Detailed 3D structural information obtained by this method can provide a foundation for structure-based drug design
Structure-based drug design of novel antibiotics:
The high resolution 3D structure of a bacterial target or host:pathogen protein-protein complex is an invaluable resource for rational drug design of novel anti-infectives. Computational methods such as high-throughput virtual screening are combined with X-ray crystallography and fragment-based screening to generate novel drug leads.
Publications:

T. Hogg and R. Hilgenfeld:
Lead Search and Optimization: Protein Crystallography in Drug Discovery.
Comprehensive Medicinal Chemistry II (J. Taylor & D. Triggle, eds.), Elsevier, Amsterdam (2007)

J. Deiwick, S.P. Salcedo, E. Boucrot, S.M. Gilliland, T. Henry, N. Petermann, S.R. Waterman, J.P. Gorvel, D.W. Holden & S. Meresse:
The translocated Salmonella effector proteins SseF and SseG interact and are required to establish an intracellular replication niche.
Infect. Immun. 74, 6965-6972 (2006).

Chakravortty D., M. Rohde, L. Jäger, J. Deiwick, and M. Hensel. 2005. Formation of a novel surface structure encoded by Salmonella Pathogenicity Island 2., EMBO J 24:2043–52.

Hogg, T., U. Mechold, H. Malke, M. Cashel, and R. Hilgenfeld, 2004. Conformational antagonism between opposing active sites in a bifunctional RelA/SpoT homolog modulates (p)ppGpp metabolism during the stringent response. Cell 117:57-68.

Deiwick J., Rappl C., Stender S., Jungblut P. R. ,Hensel M. (2004) From gene regulation to a map of Salmonella Pathogenicity Island 2-encoded proteins, Bio-Forum International (2004)

Mesters, J.R., Hogg, T. and Hilgenfeld, R. (2003). G proteins. In: Encyclopedia of Life Sciences, Nature Publishing Group, Macmillan Publishers, London.

Rappl, C., J. Deiwick, and M. Hensel .2003. Acidic pH is required for the functional assembly of the type III secretion system encoded by Salmonella pathogenicity island 2. FEMS Microbiology Letters 226:363-72

Hogg, T., Mesters, J.R. and Hilgenfeld, R. (2002). Inhibitory mechanisms of antibiotics targeting elongation factor Tu. Current Protein & Peptide Science 3, 121-131.

Deiwick, J., C. Rappl, S. Stender, P. R. Jungblut, and M. Hensel. 2002. Proteomic approaches to Salmonella Pathogenicity Island 2 encoded proteins and the SsrAB regulon.. Proteomics 2:792-9.

Hogg, T., Smatanova, I.K., Bezouska, K., Ulbrich, N. and Hilgenfeld, R. (2002). Sugar-mediated lattice contacts in crystals of a plant glycoprotein. Acta Cryst. D58, 1734-1739.

Nikolaus, T., Deiwick, J., Rappl, C., Freeman, J. A., Schroder, W., Miller, S. I., Hensel, M., (2001) SseBCD Proteins are secreted by the Type III Secretion System of Salmonella Pathogenicity Island 2 and function as a translocon J. Bacteriol., 183, 6036-6045.

Rau, A., Hogg, T., Marquardt, R. and Hilgenfeld, R. (2001). A new lysozyme fold: Structure of the muramidase from Streptomyces coelicolor at 1.65 Å resolution. J. Biol. Chem. 276, 31994-31999.

Deiwick J., Rappl C., Stender S., Jungblut P. R. ,Hensel M. (2001) Proteomic approaches to Salmonella Pathogenicity Island 2-encoded proteins and the SsrAB regulon (2001) Proteomics, 2, 792-799.

Hilgenfeld, R., Mesters, J.R. and Hogg, T. (2000). Insights into the GTPase mechanism of EF-Tu from structural studies. In: The Ribosome: Structure, Function, Antibiotics and Cellular Interactions, ASM Press, Washington, D.C., pp. 347-357.

Beuzon, C.R., Banks, G. Deiwick, J., Hensel, M., und Holden, D.W. (1999) pH-dependent secretion of SseB, a product of the SPI-2 type III secretion system of Salmonella typhimurium. Mol.Microbiol. 33, 806-816.

Deiwick, J., Nikolaus, T., Ergodan, S. und Hensel, M. (1999) Environmental regulation of Salmonella Pathogenicity Island 2 gene expression. Mol.Microbiol. 31, 1759-1774.

Deiwick, J. und Hensel, M. (1999) Regulation of virulence genes by environmental signals in Salmonella typhimurium. Electrophoresis 20, 813-819.

Deiwick, J., Nikolaus, T., Shea, J.E., Gleeson, C., Holden, D.W. und Hensel, M. (1998) Mutations in Salmonella Pathogenicity Island 2 (SPI2) genes affecting transcription of SPI1 genes und resistance to antimicrobial agents. J.Bacteriol. 180, 4775-4780.