title: New aspects on the mechanism of GroEL-assisted protein folding.
abstract: The mechanism of assisted protein folding by the chaperonin GroEL alone or in complex with the co-chaperonin GroES and in the presence or absence of nucleotides has been subject to extensive investigations during the last years. In this paper we present data where we have inactivated GroEL by stepwise blocking the nucleotide binding sites using the non-hydrolyzable ATP analogue, (Cr(H2O)4)3+ATP. We correlated the amount of accessible nucleotide binding sites with the residual ATP hydrolysis activity of GroEL as well as the residual refolding activity for two different model substrates. Under the conditions used, folding of the substrate proteins and ATP hydrolysis were directly proportional to the residual, accessible nucleotide binding sites. In the presence of GroES, 50% of the nucleotide binding sites were protected from inactivation by CrATP and the resulting protein retains 50% of both ATPase and refolding activity. The results strongly suggest that under the conditions used in our experiments, the nucleotide binding sites are additive in character and that by blocking of a certain number of binding sites a proportional amount of ATP hydrolysis and refolding activities are inactivated. The experiments including GroES suggest that full catalytic activity of GroEL requires both rings of the chaperonin. Blocking of the nucleotide binding sites of one ring still allows function of the second ring.
authors: Fachbereich Chemie der Universität Kaiserslautern, Erwin Schrödinger Strasse, 67663, Kaiserslautern, Germany.
authors: Petra
authors: P
authors: Guhr
authors: Sonja
authors: S
authors: Neuhofen
authors: Carol
authors: C
authors: Coan
authors: John G
authors: JG
authors: Wise
authors: Pia D
authors: PD
authors: Vogel
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D001665:Binding Sites; D018834:Chaperonin 60 / Q000037:antagonists & inhibitors / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy; D017510:Protein Folding; D013997:Time Factors
chemical_list: D018834:Chaperonin 60; D000255:Adenosine Triphosphate