title: Kinetics of folding and unfolding of beta beta-tropomyosin.
abstract: The kinetics of folding from random coils to two-chain coiled coils of beta beta-tropomyosin was studied by stopped-flow CD (SFCD) in the backbone region (222 nm). Two species were studied: the reduced form and the doubly disulfide cross-linked form. The proteins were totally unfolded in 6M urea-saline buffer, then refolded by tenfold dilution into benign buffer. In the refolding medium, they spontaneously recover the two-chain coiled-coil structure. Reduced beta beta refolds in at least two stages: one or more fast phases (< 0.04 s), in which an intermediate with 71% of the equilibrium ellipticity forms, followed by a slower time-resolvable phase that completes the folding. The slow phase is first order, signifying that dimerization occurs in the fast phase. The time constant of the slow phase is 2 s at 20 degrees C and requires activation parameters of delta S not equal to = -7 +/- 0.3 cal/mol.K, delta H not equal to = 15 +/- 1 kcal/mol. These results are very similar to those previously found for the reduced genetic variant alpha alpha-tropomyosin. In contrast, refolding of doubly disulfide cross-linked beta beta is complete within the dead time (< 0.04 s), whereas the singly cross-linked alpha alpha species also displays a slow phase. The opposite process, unfolding reduced beta beta from the coiled-coil state, is complete within the dead time, as in the alpha alpha variant.
authors.affiliation: Department of Chemistry, Washington University, St. Louis, Missouri 63130.
authors.forename: J
authors.initials: J
authors.lastname: Mo
authors.forename: M E
authors.initials: ME
authors.lastname: Holtzer
authors.forename: A
authors.initials: A
authors.lastname: Holtzer
mesh_terms: D000818:Animals; D007700:Kinetics; D046911:Macromolecular Substances; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011817:Rabbits; D014335:Tropomyosin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D046911:Macromolecular Substances; D014335:Tropomyosin