Monomer topology defines folding speed of heptamer.
Small monomeric proteins often fold in apparent two-state processes with folding speeds dictated by their native-state topology. Here we test, for the first time, the influence of monomer topology on the folding speed of an oligomeric protein: the heptameric cochaperonin protein 10 (cpn10), which in the native state has seven beta-barrel subunits noncovalently assembled through beta-strand pairing. Cpn10 is a particularly useful model because equilibrium-unfolding experiments have revealed that the denatured state in urea is that of a nonnative heptamer. Surprisingly, refolding of the nonnative cpn10 heptamer is a simple two-state kinetic process with a folding-rate constant in water (2.1 sec(-1); pH 7.0, 20 degrees C) that is in excellent agreement with the prediction based on the native-state topology of the cpn10 monomer. Thus, the monomers appear to fold as independent units, with a speed that correlates with topology, although the C and N termini are trapped in beta-strand pairing with neighboring subunits. In contrast, refolding of unfolded cpn10 monomers is dominated by a slow association step.
Bascos
Neil
N
Molecular and Cellular Biology Graduate Program, Tulane University, New Orleans, Louisiana 70112, USA.
Guidry
Jesse
J
Wittung-Stafshede
Pernilla
P
D018835:Chaperonin 10 / Q000032:analysis / Q000737:chemistry*; D005453:Fluorescence; D019791:Guanidine / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D017510:Protein Folding; D014508:Urea / Q000737:chemistry
D018835:Chaperonin 10; D014508:Urea; D019791:Guanidine