HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: M400048-MCP200v1 3/8/809    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Glossary Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Terradot, L. Articles by Waksman, G. Search for Related Content PubMed PubMed Citation Articles by Terradot, L. Articles by Waksman, G. Social Bookmarking                      What's this?

Molecular & Cellular Proteomics 3:809-819, 2004.
© 2004 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Research

Biochemical Characterization of Protein Complexes from the Helicobacter pylori Protein Interaction Map

Strategies for Complex Formation and Evidence for Novel Interactions Within Type IV Secretion Systems^*

Laurent Terradot^,, Nathan Durnell, Min Li, Ming Li, Jeremiah Ory, Agnes Labigne^¶, Pierre Legrain^||,**, Frederic Colland^|| and Gabriel Waksman^,,,

From the Washington University School of Medicine, Department of Biochemistry and Molecular Biophysics, 660 South Euclid Avenue, Campus Box 8231, St. Louis, MO 63110; Institute of Structural Molecular Biology, Birkbeck College and University College London, University of London, Malet Street, Bloomsbury, London WC1E 7HX, United Kingdom; ^¶ Unité de Pathogénie Bactérienne des Muqueuses, Institut Pasteur, 75724 Paris Cedex 15, France; ^|| Hybrigenics, 3-5 Impasse Reille, 75014 Paris, France; and University College London, Department of Biochemistry and Molecular Biology, Gower Street, London WC1E 6BT, United Kingdom

We have investigated a large set of interactions from the Helicobacter pylori protein interaction map previously identified by high-throughput yeast two-hybrid (htY2H)-based methods. This study had two aims: i) to validate htY2H as a source of protein-protein interaction complexes for high-throughput biochemical and structural studies of the H. pylori interactome; and ii) to validate biochemically interactions shown by htY2H to involve components of the H. pylori type IV secretion systems. Thus, 17 interactions involving 31 proteins and protein fragments were studied, and a general strategy was designed to produce protein-interacting partners for biochemical and structural characterization. We show that htY2H is a valid source of protein-protein complexes for high-throughput proteome-scale characterization of the H. pylori interactome, because 76% of the interactions tested were confirmed biochemically. Of the interactions involving type IV secretion proteins, three could be confirmed. One interaction is between two components of the type IV secretion apparatus, ComB10 and ComB4, which are VirB10 and VirB4 homologs, respectively. Another interaction is between a type IV component (HP0525, a VirB11 homolog) and a non-type IV secretion protein (HP01451), indicating that proteins other than the core VirB (1-11)-VirD4 proteins may play a role in type IV secretion. Finally, a third interaction was biochemically confirmed between CagA, a virulence factor secreted by the type IV secretion system encoded by the Cag pathogenicity island, and a non-type IV secretion protein, HP0496.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				S. Kutter, R. Buhrdorf, J. Haas, W. Schneider-Brachert, R. Haas, and W. Fischer Protein Subassemblies of the Helicobacter pylori Cag Type IV Secretion System Revealed by Localization and Interaction Studies J. Bacteriol., March 15, 2008; 190(6): 2161 - 2171. [Abstract] [Full Text] [PDF]

				M. Y. Abajy, J. Kopec, K. Schiwon, M. Burzynski, M. Doring, C. Bohn, and E. Grohmann A Type IV-Secretion-Like System Is Required for Conjugative DNA Transport of Broad-Host-Range Plasmid pIP501 in Gram-Positive Bacteria J. Bacteriol., March 15, 2007; 189(6): 2487 - 2496. [Abstract] [Full Text] [PDF]

				M. R. Couturier, E. Tasca, C. Montecucco, and M. Stein Interaction with CagF Is Required for Translocation of CagA into the Host via the Helicobacter pylori Type IV Secretion System Infect. Immun., January 1, 2006; 74(1): 273 - 281. [Abstract] [Full Text] [PDF]

				M. E. Cusick, N. Klitgord, M. Vidal, and D. E. Hill Interactome: gateway into systems biology Hum. Mol. Genet., October 15, 2005; 14(suppl_2): R171 - R181. [Abstract] [Full Text] [PDF]

				E. Formstecher, S. Aresta, V. Collura, A. Hamburger, A. Meil, A. Trehin, C. Reverdy, V. Betin, S. Maire, C. Brun, et al. Protein interaction mapping: A Drosophila case study Genome Res., March 1, 2005; 15(3): 376 - 384. [Abstract] [Full Text] [PDF]