title: Kinetics of folding of alpha alpha-tropomyosin subsequences.
abstract: The kinetics of folding random coils of alpha alpha-tropomyson (Tm) subsequences to two-chain coiled coils was studied by stopped-flow CD. Subsequences studied were those comprising residues 11-127 (11Tm127), 142-281 (142Tm281), 1-189 (1Tm189), and 190-284 (190Tm284) of the parent 284-residue alpha-tropomyosin chain. Unlike the parent, subsequences 1Tm189 and 11Tm127 fold within the dead time of the instrument (less than 0.04 s). Like the parent, subsequences 142Tm281 and 190Tm284 fold in two phases. In the fast phase, 45% and 32%, respectively, of the equilibrium helical content form. In the time-resolvable, first-order slow phase (k-1 = 2.7 s at 20 degrees C for 142Tm281 and k-1 = 2.0 s at 15 degrees C for 190Tm284), the remaining structure forms. Neither reduced 142Tm281 nor 190Tm284 show any dependence of the rate on concentration, so chain association occurs in the fast phase. Like the parent 142Tm281 forms more helical content in the fast phase when cross-linked at C-190, and the remaining structure forms slowly with rate parameters similar to those of the reduced species. Comparison of the folding behavior of C- and N-terminal subsequences with that of the parent protein suggests that the slow phase in the parent is caused by a folding bottleneck somewhere nearer the C-terminus. However, rapid association and partial folding near the N-terminus is not necessary for prompt folding, since even 190Tm284 chains associate and partially fold very rapidly (less than 0.04 s), and then complete the folding in seconds.
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; D002942:Circular Dichroism; D007700:Kinetics; D009206:Myocardium / Q000378:metabolism; D011487:Protein Conformation; D011817:Rabbits; D014335:Tropomyosin / Q000737:chemistry*
chemical_list: D014335:Tropomyosin