Hepatitis C Virus NS5A Protein Is a Substrate for the Peptidyl-prolylcis/transIsomerase Activity of Cyclophilins A and B

Autor: Ralf Bartenschlager, Xavier Hanoulle, Guy Lippens, Aurélie Badillo, Jean-Michel Wieruszeski, François Penin, Isabelle Landrieu, Dries Verdegem
Přispěvatelé: Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), 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), Department of Molecular Virology (DMV), Universität Heidelberg [Heidelberg], Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Universität Heidelberg [Heidelberg] = Heidelberg University
Rok vydání: 2009
Předmět:
Circular dichroism
viruses
Cypa
Hepacivirus
Isomerase
Viral Nonstructural Proteins
Biochemistry
Cyclophilins
03 medical and health sciences
Protein structure
Cyclosporin a
Humans
Peptidyl-prolyl cis-trans isomerase activity
Nuclear Magnetic Resonance
Biomolecular
Molecular Biology
030304 developmental biology
0303 health sciences
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Structural Biology [q-bio.BM]
biology
030306 microbiology
Active site
virus diseases
Cell Biology
biochemical phenomena
metabolism
and nutrition
biology.organism_classification
Recombinant Proteins
digestive system diseases
Protein Structure
Tertiary
3. Good health
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biomolecules [q-bio.BM]
Mutation
Protein Structure and Folding
biology.protein
Cyclophilin A
Two-dimensional nuclear magnetic resonance spectroscopy
Protein Binding
Zdroj: Journal of Biological Chemistry
Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2009, epub ahead of print. ⟨10.1074/jbc.M809244200⟩
Journal of Biological Chemistry, 2009, epub ahead of print. ⟨10.1074/jbc.M809244200⟩
ISSN: 1083-351X
0021-9258
DOI: 10.1074/jbc.m809244200
Popis: International audience; We report a biochemical and structural characterization of domain 2 of the non-structural 5A protein (NS5A) from the JFH1 Hepatitis C virus strain and its interactions with Cyclophilins A and B (CypA and CypB). Gel filtration chromatography, circular dichroism spectroscopy and finally NMR spectroscopy all indicate the natively unfolded nature of this NS5A-D2 domain. Because mutations in this domain have been linked to Cyclosporin A (CsA) resistance, we used NMR spectroscopy to investigate potential interactions between NS5A-D2 and cellular CypA and CypB. We observed a direct molecular interaction between NS5A-D2 and both Cyclophilins. The interaction surface on the Cyclophilins corresponds to their active site, whereas on NS5A-D2, it proved distributed over the many proline residues of the domain. NMR heteronuclear exchange spectroscopy yielded direct evidence that many proline residues in NS5A-D2 form a valid substrate for the enzymatic peptidyl-prolyl cis/trans isomerase (PPIase) activity of CypA and CypB.
Databáze: OpenAIRE
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