Folding and unfolding of γTIM monomers and dimers

Abstract

Kinetic simulations of the folding and unfolding of triosephosphate isomerase (TIM) from yeast were conducted using a single monomer γTIM polypeptide chain that folds as a monomer and two γTIM chains that fold to the native dimer structure. The basic protein model used was a minimalist Gō model using the native structure to determine attractive energies in the protein chain. For each simulation type - monomer unfolding, monomer refolding, dimer unfolding, and dimer refolding - thirty simulations were conducted, successfully capturing each reaction in full. Analysis of the simulations demonstrates four main conclusions. First, all four simulation types have a similar "folding order", i.e., they have similar structures in intermediate stages of folding between the unfolded and folded state. Second, despite this similarity, different intermediate stages are more or less populated in the four different simulations, with 1), no intermediates populated in monomer unfolding; 2), two intermediates populated with β2- β4 and β1-β5 regions folded in monomer refolding; 3), two intermediates populated with β2- β3 and β2-β4 regions folded in dimer unfolding; and 4), two intermediates populated with β1- β5 and β1-β5 + β6 + β7 + β8 regions folded in dimer refolding. Third, simulations demonstrate that dimer binding and unbinding can occur early in the folding process before complete monomer-chain folding. Fourth, excellent agreement is found between the simulations and MPAX (misincorporation proton alkyl exchange) experiments. In total, this agreement demonstrates that the computational Gō model is accurate for γTIM and that the energy landscape of γTIM appears funneled to the native state. © 2007 by the Biophysical Society.