title: Asymmetric photocross-linking of singly phosphorylated smooth muscle heavy meromyosin.
abstract: Although activities of smooth muscle myosin are regulated by phosphorylation, the molecular mechanisms of regulation have not been fully established. Phosphorylation of both heads of myosin is known to activate ATPase and motor activities, but the effects of phosphorylation of only one of the heads have not been established. Such information on singly phosphorylated myosin can serve to elucidate the molecular mechanism of the phosphorylation-dependent regulation. To understand the structural properties of the singly phosphorylated state, we prepared singly phosphorylated heavy meromyosin (HMM) containing a photoreactive benzophenone-labeled RLC and examined its photocross-linking reactivity. The two heads in the singly phosphorylated HMM showed different reactivities. The dephosphorylated RLC in the singly phosphorylated HMM was cross-linked to a heavy chain, like that in the dephosphorylated HMM, whereas the phosphorylated RLC did not react, like that in the fully phosphorylated HMM. These results indicate that the two heads of the singly phosphorylated HMM have an asymmetric structure, suggesting that phosphorylation of one head can to some extent activate smooth muscle HMM.
authors.affiliation: Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Takeya
authors.forename: Masayuki
authors.initials: M
authors.lastname: Takahashi
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Katoh
authors.forename: Michio
authors.initials: M
authors.lastname: Yazawa
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001577:Benzophenones / Q000737:chemistry / Q000378:metabolism; D002645:Chickens; D003432:Cross-Linking Reagents / Q000378:metabolism*; D008027:Light; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000378:metabolism*; D015879:Myosin Subfragments / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001577:Benzophenones; D003432:Cross-Linking Reagents; D015879:Myosin Subfragments; D020033:Protein Isoforms; C047723:benzophenone