Kinetic partitioning. Poising SecB to favor association with a rapidly folding ligand.
Chaperones are a class of proteins that possess the remarkable ability to selectively bind polypeptides that are in a nonnative state. The selectivity of SecB, a molecular chaperone in Escherichia coli, for its ligands can be explained in part by a kinetic partitioning between folding of the polypeptide and association with SecB. It has clearly been established that kinetic partitioning can be poised to favor association with SecB by changing the rate constant for folding of the ligand. We now demonstrate that binding to SecB can be given a kinetic advantage over the pathway for folding by modulating the properties of the chaperone. By poising SecB to expose a hydrophobic patch, we were able to detect a complex between SecB and maltose-binding protein under conditions in which rapid folding of the polypeptide otherwise precludes formation of a kinetically stable complex. The data presented here are interpreted within the framework of a kinetic partitioning between binding to SecB and folding of the polypeptide. We propose that exposure of a hydrophobic patch on SecB increases the surface area for binding and thereby increases the rate constant for association. In this way association of SecB with the polypeptide ligand has a kinetic advantage over the pathway for folding.
Diamond
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Department of Biochemistry and Biophysics, Washington State University, Pullman, Washington 99164-4660, USA. diamond@membrane.chem.wsu.edu
Randall
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LL
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins / Q000378:metabolism*; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism; D029968:Escherichia coli Proteins; D007700:Kinetics; D008024:Ligands; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D010455:Peptides / Q000378:metabolism; D017510:Protein Folding; D013696:Temperature
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D008024:Ligands; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D010455:Peptides; C057842:SecB protein, Bacteria; C461599:maltose transport system, E coli