title: Probing the energy landscape of protein folding/unfolding transition states.
abstract: Previous molecular dynamics (MD) simulations of the thermal denaturation of chymotrypsin inhibitor 2 (CI2) have provided atomic-resolution models of the transition state ensemble that is well supported by experimental studies. Here, we use simulations to further investigate the energy landscape around the transition state region. Nine structures within approximately 35 ps and 3 A C(alpha) RMSD of the transition state ensemble identified in a previous 498 K thermal denaturation simulation were quenched under the quasi-native conditions of 335 K and neutral pH. All of the structures underwent hydrophobically driven collapse in response to the drop in temperature. Structures less denatured than the transition state became structurally more native-like, while structures that were more denatured than the transition state tended to show additional loss of native structure. The structures in the immediate region of the transition state fluctuated between becoming more and less native-like. All of the starting structures had the same native-like topology and were quite similar (within 3.5 A C(alpha) RMSD). That the structures all shared native-like topology, yet diverged into either more or less native-like structures depending on which side of the transition state they occupied on the unfolding trajectory, indicates that topology alone does not dictate protein folding. Instead, our results suggest that a detailed interplay of packing interactions and interactions with water determine whether a partially denatured protein will become more native-like under refolding conditions.
authors.affiliation: Department of Medical Chemistry H165, School of Pharmacy, University of Washington, P.O. Box 357610, Seattle, WA 98195-7610, USA.
authors.forename: Deborah
authors.initials: D
authors.lastname: De Jong
authors.forename: Robert
authors.initials: R
authors.lastname: Riley
authors.forename: Darwin O V
authors.initials: DO
authors.lastname: Alonso
authors.forename: Valerie
authors.initials: V
authors.lastname: Daggett
mesh_terms: D000818:Animals; D003198:Computer Simulation; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D011498:Protein Precursors / Q000737:chemistry* / Q000378:metabolism*; D020673:Protein Renaturation; D013552:Swine; D013696:Temperature; D013816:Thermodynamics; D013997:Time Factors; D014364:Tryptophan / Q000737:chemistry; D014867:Water / Q000737:chemistry; D054804:Cathelicidins
chemical_list: D011498:Protein Precursors; D014364:Tryptophan; D014867:Water; C084994:NPG4 protein, Sus scrofa; D054804:Cathelicidins