Etudes structurale et fonctionnelle de protéines impliquées dans la virulence chez S. pneumoniae et P. aeruginosa

Abstract : This manuscript is made up of two parts The first part describes the structural study of RrgA from Streptococcus pneumoniae. This protein is a pilus-associated adhesin that is able to bind to several components of the Extra Cellular Matrix and thus, participates in the first steps of host colonization. We solved the structure of RrgA to 1.9 Å by X-Ray crystallography. We showed that RrgA folds into an elongated 4-domain structure, and these domains display both eukaryotic and prokaryotic origins. Actually, three out of the four domains are reminiscent of IgG and Cna-B structures and act like stalks to orient and display the large Integrin-like domain. We confirmed the presence of two isopeptide bonds by mass spectrometry and hypothesised that the two inserted arms in the integrin domain could explain the wide variety of substrates RrgA can bind. The second part of this manuscript focuses on the structural studies of the ATPase complex as well as ExsB, the putative pilotin of the type III secretion system from Pseudomonas aeruginosa. This bacterium is a major threat in hospital-acquired infections and the main pathogen found in cystic-fibrosis suffering patients. For the first time we were able to express and purify the ATPase PscN in complex with its partner PscL. Crystallization trials led to a very promising condition that is being refined. Moreover, we confirmed expression of the lipoprotein ExsB in P. aeruginosa that we localised in the outer membrane. To have a better understanding of this protein, we also solved its high-resolution structure that displays a novel fold and our study paves the way for coming studies concerning pilotins.
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Submitted on : Monday, November 7, 2011 - 12:10:34 PM
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  • HAL Id : tel-00638763, version 1



Thierry Izoré. Etudes structurale et fonctionnelle de protéines impliquées dans la virulence chez S. pneumoniae et P. aeruginosa. Biologie structurale [q-bio.BM]. Université de Grenoble, 2011. Français. ⟨NNT : 2011GRENV047⟩. ⟨tel-00638763⟩



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