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Radicicol-sensitive peptide binding to the N-terminal portion of GRP94
Journal article   Open access   Peer reviewed

Radicicol-sensitive peptide binding to the N-terminal portion of GRP94

Shawn Vogen, Tali Gidalevitz, Chhanda Biswas, Birgitte B Simen, Eytan Stein, Funda Gulmen and Yair Argon
The Journal of biological chemistry, v 277(43), pp 40742-40750
25 Oct 2002
PMID: 12189140
url
http://www.jbc.org/content/277/43/40742.full.pdfView
Published, Version of Record (VoR) Open
url
https://doi.org/10.1074/jbc.M205323200View
Published, Version of Record (VoR) Open

Abstract

Amino Acid Sequence Animals Binding Sites HSP70 Heat-Shock Proteins - chemistry HSP70 Heat-Shock Proteins - metabolism Lactones - pharmacology Ligands Macrolides Membrane Proteins - chemistry Membrane Proteins - metabolism Mice Molecular Sequence Data Peptides - metabolism Protein Binding Recombinant Proteins - chemistry Recombinant Proteins - metabolism
GRP94 is a molecular chaperone that carries immunologically relevant peptides from cell to cell, transferring them to major histocompatibility proteins for presentation to T cells. Here we examine the binding of several peptides to recombinant GRP94 and study the regulation and site of peptide binding. We show that GRP94 contains a peptide-binding site in its N-terminal 355 amino acids. A number of peptides bind to this site with low on- and off-rates and with specificity that is distinct from that of another endoplasmic reticulum chaperone, BiP/GRP78. Binding to the N-terminal fragment is sufficient to account for the peptide binding activity of the entire molecule. Peptide binding is inhibited by radicicol, a known inhibitor of the chaperone activities of HSP90-family proteins. However, the peptide-binding site is distinct from the radicicol-binding pocket, because both can bind to the N-terminal fragment simultaneously. Furthermore, peptide binding does not cause the same conformational change as does binding of radicicol. When the latter binds to the N-terminal domain, it induces a conformational change in the downstream, acidic domain of GRP94, as measured by altered gel mobility and loss of an antibody epitope. These results relate the peptide-binding activity of GRP94 to its other function as a chaperone.

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Web of Science research areas
Biochemistry & Molecular Biology
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