title: The chaperone cofactor Hop/p60 interacts with the cytosolic chaperonin-containing TCP-1 and affects its nucleotide exchange and protein folding activities.
abstract: The folding of protein structures often requires the presence of molecular chaperones and/or chaperonin complexes. We here investigated the inhibitory effects of the chaperone cofactors Hop/p60 and Hap46. By coimmunoprecipitation, we observed a direct interaction of the eukaryotic chaperonin-containing TCP-1 (CCT) purified from rabbit reticulocyte lysate with Hop/p60. By contrast, Hap46 was not coprecipitated. Binding of Hop/p60 to CCT is dependent on the presence of ATP or ADP and occurs through carboxyl-terminal sequences of Hop/p60. Hop/p60 significantly stimulates nucleotide exchange on CCT but not its ATPase activity, while Hap46 has no effects. We used denatured firefly luciferase as a model protein and found decreased binding to CCT in the presence of Hop/p60 and ATP. This coincides with the inhibitory effect of Hop/p60 on luciferase reactivation in an assay using purified CCT in combination with hsc70 and hsp40. We also observed that an antibody directed against one of the subunits of CCT efficiently inhibits refolding in a system which depends on crude reticulocyte lysate.
authors.affiliation: Universität Heidelberg, Biochemie-Zentrum Heidelberg, Biologische Chemie, Im Neuenheimer Feld 501, D-69120 Heidelberg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Gebauer
authors.forename: R
authors.initials: R
authors.lastname: Melki
authors.forename: U
authors.initials: U
authors.lastname: Gehring
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins / Q000378:metabolism*; D001517:Coleoptera / Q000201:enzymology; D003600:Cytosol / Q000378:metabolism; D006358:Hot Temperature; D008156:Luciferases / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011489:Protein Denaturation; D017510:Protein Folding; D011817:Rabbits
chemical_list: D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D008156:Luciferases; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins