Hepatitis C Virus NS5A Protein Is a Substrate for the Peptidyl-prolyl cis/trans Isomerase Activity of Cyclophilins A and B

被引：135

作者：

Hanoulle, Xavier ^{[1
]}

Badillo, Aurelie ^{[2
]}

Wieruszeski, Jean-Michel ^{[1
]}

Verdegem, Dries ^{[1
]}

Landrieu, Isabelle ^{[1
]}

Bartenschlager, Ralf ^{[3
]}

Penin, Francois ^{[2
]}

Lippens, Guy ^{[1
]}

机构：

[1] Univ Sci & Technol Lille, CNRS, IFR 147, Unite Glycobiol Struct & Fonctionnelle,UMR 8576, F-59655 Villeneuve Dascq, France

[2] Univ Lyon, Inst Biol & Chim Prot, UMR 5086, CNRS,IFR 128, F-69397 Lyon, France

[3] Heidelberg Univ, Dept Mol Virol, D-69120 Heidelberg, Germany

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 2009年 / 284卷 / 20期

关键词：

IMMUNODEFICIENCY-VIRUS; NONSTRUCTURAL PROTEIN; CYCLOSPORINE-A; BINDING-PROTEIN; REPLICATION; SENSITIVITY; RESISTANCE; CATALYSIS; DOMAIN; IDENTIFICATION;

D O I：

10.1074/jbc.M809244200

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We report here a biochemical and structural characterization of domain 2 of the nonstructural 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 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 to be 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.

引用

页码：13589 / 13601

页数：13

共 71 条

[11] DETERMINATION OF HELIX AND BETA-FORM OF PROTEINS IN AQUEOUS-SOLUTION BY CIRCULAR-DICHROISM [J].