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ETUDES STRUCTURALES DES PROTEINES PAR SPECTROMETRIE DE MASSE COUPLEE AUX ECHANGES HYDROGENE/DEUTERIUM ET A LA RETICULATION CHIMIQUE

Abstract : Proteins are involved in many biological processes. They might be targets for medical treatments as well as therapeutic agents. A detailed knowledge of protein structure and a characterization of protein complexes are important to understand protein functions in a cell. In this study, we developed two new methods, which use mass spectrometry, to elucidate protein structure. These methods were then applied with success on a biological study. We improved a method that combines H/D exchange experiments with mass spectrometry on a model protein: PBP-2X. We show that the combination of three proteases increases the sequence coverage of the protein and the spatial resolution in the determination of interest areas. We developed a method, which associates intramolecular cross-linking and mass spectrometry on a model protein: the cytochrome c. Distance constraints determined by this way will be included in a bioinformatic project, that could give the folding family of a protein of which the tri-dimensional structure is unknown. We applied these two methods on proteins which are involved in type III protein secretion system from Pseudomonas aeruginosa: PcrV and PcrG. Experimental data (accessibility, secondary structures and distance constraints) are in agreement with the structural predictions on PcrV. PcrG is mainly unstructured. PcrG and PcrV are in interaction through their coiled-coil domains. Complexation between the two proteins induces conformational changes on PcrV, which could stabilize PcrG
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https://tel.archives-ouvertes.fr/tel-00009698
Contributor : Laetitia Cravello <>
Submitted on : Thursday, July 7, 2005 - 9:12:56 AM
Last modification on : Friday, November 6, 2020 - 3:57:09 AM
Long-term archiving on: : Friday, April 2, 2010 - 10:04:36 PM

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  • HAL Id : tel-00009698, version 1

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Laetitia Cravello. ETUDES STRUCTURALES DES PROTEINES PAR SPECTROMETRIE DE MASSE COUPLEE AUX ECHANGES HYDROGENE/DEUTERIUM ET A LA RETICULATION CHIMIQUE. Sciences pharmaceutiques. Université Joseph-Fourier - Grenoble I, 2005. Français. ⟨tel-00009698⟩

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