title: Kinetics of structural changes in the relay loop and SH3 domain of myosin.
abstract: The intrinsic fluorescence of smooth muscle myosin signals conformational changes associated with different catalytic states of the ATPase cycle. To elucidate this relationship, we have examined the pre-steady-state kinetics of nucleotide binding, hydrolysis, and product release in motor domain-essential light chain mutants containing a single endogenous tryptophan, either residue 512 in the rigid relay loop or residue 29 adjacent to the SH3 domain. The intrinsic fluorescence of W512 is sensitive to both nucleotide binding and hydrolysis, and appears to report structural changes at the active site, presumably through a direct connection with switch II. The intrinsic fluorescence of W29 is sensitive to nucleotide binding but not hydrolysis, and does not appear to be tightly linked with structural changes occurring at the active site. We propose that the SH3 domain may be sensitive to conformational changes in the lever arm through contacts with the essential light chain.
authors.affiliation: Department of Molecular Physiology and Biophysics, College of Medicine, University of Vermont, Burlington, VT 05405-0075, USA.
authors.forename: Marilyn
authors.initials: M
authors.lastname: van Duffelen
authors.forename: Lynn R
authors.initials: LR
authors.lastname: Chrin
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Berger
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry*; D019943:Amino Acid Substitution; D001665:Binding Sites; D017301:Ca(2+) Mg(2+)-ATPase / Q000737:chemistry*; D006868:Hydrolysis; D007700:Kinetics; D020409:Molecular Motor Proteins / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D024745:Smooth Muscle Myosins / Q000737:chemistry*; D013329:Structure-Activity Relationship; D014364:Tryptophan / Q000737:chemistry*; D018909:src Homology Domains
chemical_list: D020409:Molecular Motor Proteins; D014364:Tryptophan; D000255:Adenosine Triphosphate; D017301:Ca(2+) Mg(2+)-ATPase; D024745:Smooth Muscle Myosins