Specificity of peptide-induced depolymerization of the recombinant carboxy-terminal fragment of BiP/GRP78.
In the present study, we have used a non-denaturing gel electrophoresis assay to characterize the specificity of the peptide-induced depolymerization process of the isolated recombinant C-terminal domain (C30) of the molecular chaperone BiP, in the presence of specific synthetic peptides and with the neuropeptide Substance P. In the absence of peptidic ligand, C30 self-associates readily into multiple oligomeric species. Upon peptide addition, C30 oligomers convert into dimers, then into monomers. Our data indicate that the algorithm we previously developed to predict putative BiP binding sites in any protein sequence is also a good indicator as to whether a peptide can efficiently induce depolymerization of the C-terminal peptide binding domain and stimulate the ATPase activity of the full-length protein.
King
L
L
Center for Pharmaceutical Biotechnology, College of Pharmacy, Chicago, Illinois 60607-7173, USA.
Chevalier
M
M
Blond
S Y
SY
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001704:Biopolymers / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D019281:Dimerization; D000091342:Endoplasmic Reticulum Chaperone BiP; D004926:Escherichia coli / Q000235:genetics; D006360:Heat-Shock Proteins; D066298:In Vitro Techniques; D051379:Mice; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology*; D011487:Protein Conformation / Q000187:drug effects; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D001704:Biopolymers; D002352:Carrier Proteins; D017931:DNA Primers; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D018832:Molecular Chaperones; D010446:Peptide Fragments; D010455:Peptides; D011994:Recombinant Proteins