title: Im7 folding mechanism: misfolding on a path to the native state.
abstract: Many proteins populate collapsed intermediate states during folding. In order to elucidate the nature and importance of these species, we have mapped the structure of the on-pathway intermediate of the four-helix protein, Im7, together with the conformational changes it undergoes as it folds to the native state. Kinetic data for 29 Im7 point mutants show that the intermediate contains three of the four helices found in the native structure, packed around a specific hydrophobic core. However, the intermediate contains many non-native interactions; as a result, hydrophobic interactions become disrupted in the rate-limiting transition state before the final helix docks onto the developing structure. The results of this study support a hierarchical mechanism of protein folding and explain why the misfolding of Im7 occurs. The data also demonstrate that non-native interactions can play a significant role in folding, even for small proteins with simple topologies.
authors.affiliation: School of Biochemistry and Molecular Biology, University of Leeds, Mount Preston Street, Leeds, LS2 9JT, UK.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Capaldi
authors.forename: Colin
authors.initials: C
authors.lastname: Kleanthous
authors.forename: Sheena E
authors.initials: SE
authors.lastname: Radford
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D003087:Colicins; D005470:Fluorometry; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D020673:Protein Renaturation; D017433:Protein Structure, Secondary / Q000187:drug effects; D013816:Thermodynamics; D014508:Urea / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D003087:Colicins; C007104:colicin immunity proteins; D014508:Urea