title: Structural requirements for phosphorylation of myosin regulatory light chain from smooth muscle.
abstract: Site-directed and chimeric mutations of myosin regulatory light chains were used to identify residues important for phosphorylation of Ser19 by smooth muscle myosin light chain kinase. Arg16 and hydrophobic residues C-terminal of Ser19 in smooth muscle light chain were important substrate determinants in the intact protein. However, changes in the kinetic properties with mutations in the light chain were substantially smaller than results reported with structurally similar synthetic peptide substrates. These results together with the low Vmax value for short peptide substrates containing the consensus phosphorylation sequence suggest that there may be additional sites of interactions between the kinase and protein substrate. Chimeras of skeletal and smooth muscle light chains were constructed with exchanges at the N terminus and subdomains I, II, III, and IV. Analysis of results obtained on the kinetic properties for phosphorylation showed that subdomains I and II contribute to high Vmax values. Thus, a region distant from the consensus phosphorylation sequence in smooth muscle light chain is also an important substrate determinant for myosin light chain kinase.
authors.affiliation: Department of Physiology, University of Texas Southwestern Medical Center at Dallas 75235-9040.
authors.forename: G
authors.initials: G
authors.lastname: Zhi
authors.forename: B P
authors.initials: BP
authors.lastname: Herring
authors.forename: J T
authors.initials: JT
authors.lastname: Stull
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000737:chemistry; D009130:Muscle, Smooth / Q000737:chemistry*; D009154:Mutation; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D011817:Rabbits
chemical_list: D009219:Myosin-Light-Chain Kinase; D009218:Myosins