title: Characterization of oligomers as kinetic intermediates in myosin subfragment 1-induced polymerization of G-actin.
abstract: The nature of the kinetic intermediates involved in S1-induced polymerization of G-actin into F-acto-S1-decorated filaments has been investigated using as probes light scattering, the fluorescence of pyrenyl- or NBD-labeled actin, and the anisotropy of fluorescence of N-iodoacetyl-N'-(5-sulfo-1-napthyl)ethylene diamine (AEDANS)-labeled actin. With AEDANS-G-actin, the initial formation of a ternary G2S complex between two G-actin and one S1 molecules (Valentin-Ranc, C., Combeau, C., Carlier, M. F., and Pantaloni, D. (1991) J. Biol. Chem. 266, 17871-17879) has been confirmed. Data obtained with all probes are consistent with the subsequent rapid formation of G-actin-S1 oligomers in which the actin/S1 molar ratio is 2:1. Oligomers form above 0.6 microM G-actin with S1(A1) and above 3.5 microM G-actin with S1(A2), at 20 degrees C. Oligomerization is endothermic with a delta H of 14 kcal/mol. A tentative model is proposed to comprehensively account for the data and the structural features of the F-actin-S1 filament. Within this model, longitudinal actin-actin interactions take place in G2S, and lateral, hydrophobic actin-actin interactions appear upon formation of (G2S)n oligomers.
authors.affiliation: Laboratoire d'Enzymologie, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
authors.forename: C
authors.initials: C
authors.lastname: Valentin-Ranc
authors.forename: M F
authors.initials: MF
authors.lastname: Carlier
mesh_terms: D009327:4-Chloro-7-nitrobenzofurazan; D000199:Actins / Q000378:metabolism*; D000818:Animals; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D007700:Kinetics; D008027:Light; D015879:Myosin Subfragments / Q000378:metabolism*; D009282:Naphthalenesulfonates; D010446:Peptide Fragments / Q000378:metabolism; D011817:Rabbits; D012542:Scattering, Radiation; D013816:Thermodynamics
chemical_list: D000199:Actins; D005456:Fluorescent Dyes; D015879:Myosin Subfragments; D009282:Naphthalenesulfonates; D010446:Peptide Fragments; C006421:1,5-I-AEDANS; D009327:4-Chloro-7-nitrobenzofurazan