title: The myosin C-loop is an allosteric actin contact sensor in actomyosin.
abstract: Actin and myosin form the molecular motor in muscle. Myosin is the enzyme performing ATP hydrolysis under the allosteric control of actin such that actin binding initiates product release and force generation in the myosin power stroke. Non-equilibrium Monte Carlo molecular dynamics simulation of the power stroke suggested that a structured surface loop on myosin, the C-loop, is the actin contact sensor initiating actin activation of the myosin ATPase. Previous experimental work demonstrated C-loop binds actin and established the forward and reverse allosteric link between the C-loop and the myosin active site. Here, smooth muscle heavy meromyosin C-loop chimeras were constructed with skeletal (sCl) and cardiac (cCl) myosin C-loops substituted for the native sequence. In both cases, actin-activated ATPase inhibition is indicated mainly by the lower V(max). In vitro motility was also inhibited in the chimeras. Motility data were collected as a function of myosin surface density, with unregulated actin, and with skeletal and cardiac isoforms of tropomyosin-bound actin for the wild type, cCl, and sCl. Slow and fast subpopulations of myosin velocities in the wild-type species were discovered and represent geometrically unfavorable and favorable actomyosin interactions, respectively. Unfavorable interactions are detected at all surface densities tested. Favorable interactions are more probable at higher myosin surface densities. Cardiac tropomyosin-bound actin promotes the favorable actomyosin interactions by lowering the inhibiting geometrical constraint barriers with a structural effect on actin. Neither higher surface density nor cardiac tropomyosin-bound actin can accelerate motility velocity in cCl or sCl, suggesting the element initiating maximal myosin activation by actin resides in the C-loop.
authors.affiliation: Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, Minnesota 55905, USA.
authors.forename: Katalin
authors.initials: K
authors.lastname: Ajtai
authors.forename: Miriam F
authors.initials: MF
authors.lastname: Halstead
authors.forename: Miklós
authors.initials: M
authors.lastname: Nyitrai
authors.forename: Alan R
authors.initials: AR
authors.lastname: Penheiter
authors.forename: Ye
authors.initials: Y
authors.lastname: Zheng
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Burghardt
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D000205:Actomyosin / Q000737:chemistry* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000495:Allosteric Site; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009010:Monte Carlo Method; D009219:Myosin-Light-Chain Kinase / Q000737:chemistry / Q000378:metabolism; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D000199:Actins; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D000205:Actomyosin; D009219:Myosin-Light-Chain Kinase; D009218:Myosins