Molecular dissection of protein-protein interactions between integrin α5β1 and the Helicobacter pylori Cag Type IV secretion system

Autor: Lionel Ballut, Patricia Rousselle, Laurent Terradot, Rémi Fronzes, Rainer Haas, Karl Brillet, Mickael V. Cherrier, Luisa F. Jiménez-Soto, Thomas Koelblen, Célia Bergé, Junichi Takagi, Wolfgang Fischer, Roland Montserret
Přispěvatelé: Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Institut für Hygiene und Medizinische Mikrobiologie, Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Laboratoire de Biologie Tissulaire et d'ingénierie Thérapeutique UMR 5305 (LBTI), European Institute of Chemistry and Biology, Université Sciences et Technologies - Bordeaux 1, Centre de génétique moléculaire (CGM), Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Protein Conformation
[SDV]Life Sciences [q-bio]
Integrin
CHO Cells
Biology
digestive system
Biochemistry
Pilus
Microbiology
Protein–protein interaction
Type IV Secretion Systems
03 medical and health sciences
Cricetulus
Bacterial Proteins
X-Ray Diffraction
Cricetinae
Protein Interaction Mapping
Scattering
Small Angle
CagA
Animals
Humans
Secretion
[SDV.BBM]Life Sciences [q-bio]/Biochemistry
Molecular Biology
Molecular Biology
ComputingMilieux_MISCELLANEOUS
Antigens
Bacterial
030102 biochemistry & molecular biology
Helicobacter pylori
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Structural Biology [q-bio.BM]
Cell Biology
Surface Plasmon Resonance
bacterial infections and mycoses
Ligand (biochemistry)
digestive system diseases
Recombinant Proteins
Cell biology
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biomolecules [q-bio.BM]
Fibronectin
030104 developmental biology
Ectodomain
biology.protein
bacteria
Integrin alpha5beta1
Protein Binding
Zdroj: FEBS Journal
FEBS Journal, Wiley, 2017, 284 (23), pp.4143-4157. ⟨10.1111/febs.14299⟩
ISSN: 1742-464X
1742-4658
DOI: 10.1111/febs.14299⟩
Popis: The more severe strains of the bacterial human pathogen Helicobacter pylori produce a type IV secretion system (cagT4SS) to inject the oncoprotein cytotoxin-associated gene A (CagA) into gastric cells. This syringe-like molecular apparatus is prolonged by an external pilus that exploits integrins as receptors to mediate the injection of CagA. The molecular determinants of the interaction of the cagT4SS pilus with the integrin ectodomain are still poorly understood. In this study, we have used surface plasmon resonance (SPR) to generate a comprehensive analysis of the protein-protein interactions between purified CagA, CagL, CagI, CagY repeat domain II (CagYRRII ), CagY C-terminal domain (CagYB10 ) and integrin α5β1 ectodomain (α5β1E ) or headpiece domain (α5β1HP ). We found that CagI, CagA, CagL and CagYB10 but not CagYRRII were able to interact with α5β1E with affinities similar to the one observed for α5β1E interaction with its physiological ligand fibronectin. We further showed that integrin activation and its associated conformational change increased CagA, CagL and CagYB10 affinities for the receptor. Furthermore, CagI did not interact with integrin unless the receptor was in open conformation. CagI, CagA but not CagL and CagYB10 interacted with the α5β1HP . Our SPR study also revealed novel interactions between CagA and CagL, CagA and CagYB10 , and CagA and CagI. Altogether, our data map the network of interactions between host-cell α5β1 integrin and the cagT4SS proteins and suggest that activation of the receptor promotes interactions with the secretion apparatus and possibly CagA injection.
Databáze: OpenAIRE
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