title: Importance of electrostatic interactions in the rapid binding of polypeptides to GroEL.
abstract: The question of how chaperones rapidly bind non-native proteins of very different sequence and function has been examined by determining the effect of ionic strength on the refolding of barnase on GroEL, and on the thermal denaturation of barnase in the presence of GroEL and SecB. Both chaperones bind the denatured state of barnase, so lowering the T(m) value. The refolding of barnase in the presence of GroEL is multiphasic, the slowest phase corresponding to the refolding of a singly bound molecule of barnase in the complex with GroEL. The fastest phase is related to the association of barnase and GroEL. At high ratios of GroEL to barnase and low ionic strength (less than 200 mM) this fast phase corresponds to the observed rate of binding. The rate of association of barnase and GroEL was found to be highly dependent on ionic strength, and at high ionic strength (greater than 600 mM) the majority of barnase molecules escaped binding and refolded free in solution. The data are consistent with an initial, transient, ionic interaction between barnase and GroEL, before hydrophobic binding occurs, allowing diffusion-controlled association and slow dissociation of unfolded polypeptide.
authors.affiliation: MRC Unit for Protein Function and Design, University Chemical Laboratory, Cambridge, UK.
authors.forename: S
authors.initials: S
authors.lastname: Perrett
authors.forename: R
authors.initials: R
authors.lastname: Zahn
authors.forename: G
authors.initials: G
authors.lastname: Stenberg
authors.forename: A R
authors.initials: AR
authors.lastname: Fersht
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D018834:Chaperonin 60 / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008956:Models, Chemical; D009994:Osmolar Concentration; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011489:Protein Denaturation; D017510:Protein Folding; D012260:Ribonucleases / Q000737:chemistry / Q000378:metabolism*; D055672:Static Electricity
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; D010455:Peptides; C057842:SecB protein, Bacteria; D012260:Ribonucleases; C019760:Bacillus amyloliquefaciens ribonuclease