Light-triggered myosin activation for probing dynamic cellular processes.

Myosin II is an ATPase motor protein essential for many cellular functions including cell migration[1] and division.[2] In nonmuscle cells, myosin modulates protrusions at the leading edge and promotes retraction at the trailing edge during migration,[3] while during cytokinesis, myosin is required for contraction of the cleavage furrow.[4] For nonmuscle myosin, these varied functions are regulated by phosphorylation of the associated myosin regulatory light chain (mRLC) protein at Ser19, which activates the myosin complex to promote myosin assembly, contractility, and stress fiber formation.[5] Upon phosphorylation of the mRLC at both Thr18 and Ser19, these activities are further enhanced.[6 ] The dramatic effects of phosphorylation can also be recapitulated in vitro. Specifically, myosin and the proteolytic derivative heavy meromyosin (HMM),[7] which contains only one-third of the C-terminal myosin tail, exhibit low in vitro activities when associated with the nonphosphorylated mRLC. Phosphorylation of Ser19 amplifies actin-activated ATPase activities 10 – 1000-fold[8] and leads to myosin-mediated actin translocation.[9]

[1]  Miguel Vicente-Manzanares,et al.  Non-muscle myosin II takes centre stage in cell adhesion and migration , 2009, Nature Reviews Molecular Cell Biology.

[2]  G. Ellis‐Davies,et al.  Caged compounds: photorelease technology for control of cellular chemistry and physiology , 2007, Nature Methods.

[3]  B. Imperiali,et al.  Caged O-phosphorothioyl amino acids as building blocks for Fmoc-based solid phase peptide synthesis. , 2007, Tetrahedron.

[4]  B. Imperiali,et al.  Semisynthesis of unnatural amino acid mutants of paxillin: Protein probes for cell migration studies , 2007, Protein science : a publication of the Protein Society.

[5]  S. Yonemura,et al.  Regulation of myosin II dynamics by phosphorylation and dephosphorylation of its light chain in epithelial cells. , 2006, Molecular biology of the cell.

[6]  B. Imperiali,et al.  Chemical approaches for investigating phosphorylation in signal transduction networks. , 2005, Trends in cell biology.

[7]  刘金明,et al.  IL-13受体α2降低血吸虫病肉芽肿的炎症反应并延长宿主存活时间[英]/Mentink-Kane MM,Cheever AW，Thompson RW，et al//Proc Natl Acad Sci U S A , 2005 .

[8]  B. Imperiali,et al.  In Situ Photoactivation of a Caged Phosphotyrosine Peptide Derived from Focal Adhesion Kinase Temporarily Halts Lamellar Extension of Single Migrating Tumor Cells* , 2005, Journal of Biological Chemistry.

[9]  Michael E. Hahn,et al.  Photocontrol of Smad2, a multiphosphorylated cell-signaling protein, through caging of activating phosphoserines. , 2004, Angewandte Chemie.

[10]  M. Yaffe,et al.  Caged phosphopeptides reveal a temporal role for 14-3-3 in G1 arrest and S-phase checkpoint function , 2004, Nature Biotechnology.

[11]  Mitsuo Ikebe,et al.  ZIP kinase is responsible for the phosphorylation of myosin II and necessary for cell motility in mammalian fibroblasts , 2004, The Journal of cell biology.

[12]  M. Yaffe,et al.  Fluorescent caged phosphoserine peptides as probes to investigate phosphorylation-dependent protein associations. , 2003, Journal of the American Chemical Society.

[13]  Timothy J Mitchison,et al.  Dissecting Temporal and Spatial Control of Cytokinesis with a Myosin II Inhibitor , 2003, Science.

[14]  H. Hosoya,et al.  HeLa ZIP kinase induces diphosphorylation of myosin II regulatory light chain and reorganization of actin filaments in nonmuscle cells , 2001, Oncogene.

[15]  K. Trybus,et al.  Biochemical studies of myosin. , 2000, Methods.

[16]  R. Tuft,et al.  Effects of the Regulatory Light Chain Phosphorylation of Myosin II on Mitosis and Cytokinesis of Mammalian Cells* , 2000, The Journal of Biological Chemistry.

[17]  J. Sellers,et al.  Kinetics of Smooth Muscle Heavy Meromyosin with One Thiophosphorylated Head* , 2000, The Journal of Biological Chemistry.

[18]  Shuh Narumiya,et al.  Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension , 1997, Nature.

[19]  R. Cooke,et al.  In vitro actin filament sliding velocities produced by mixtures of different types of myosin. , 1997, Biophysical journal.

[20]  E. Elson,et al.  Expression of the catalytic domain of myosin light chain kinase increases paracellular permeability. , 1996, The American journal of physiology.

[21]  K. Burridge,et al.  Rho-stimulated contractility drives the formation of stress fibers and focal adhesions , 1996, The Journal of cell biology.

[22]  D. Lauffenburger,et al.  Cell Migration: A Physically Integrated Molecular Process , 1996, Cell.

[23]  T. Pollard,et al.  Phosphorylation on threonine-18 of the regulatory light chain dissociates the ATPase and motor properties of smooth muscle myosin II. , 1995, Biochemistry.

[24]  M. Ikebe,et al.  Mutagenesis of the phosphorylation site (serine 19) of smooth muscle myosin regulatory light chain and its effects on the properties of myosin. , 1994, Biochemistry.

[25]  H. Hidaka,et al.  Selective inhibition of catalytic activity of smooth muscle myosin light chain kinase. , 1987, The Journal of biological chemistry.

[26]  J. Spudich,et al.  Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination. , 1987, Science.

[27]  D. Hartshorne,et al.  Identification, phosphorylation, and dephosphorylation of a second site for myosin light chain kinase on the 20,000-dalton light chain of smooth muscle myosin. , 1986, The Journal of biological chemistry.

[28]  J. Sellers.  Mechanism of the phosphorylation-dependent regulation of smooth muscle heavy meromyosin. , 1985, The Journal of biological chemistry.

[29]  D. Hartshorne,et al.  Proteolysis of smooth muscle myosin by Staphylococcus aureus protease: preparation of heavy meromyosin and subfragment 1 with intact 20 000-dalton light chains. , 1985, Biochemistry.

[30]  J. Sellers,et al.  Reversible phosphorylation of smooth muscle myosin, heavy meromyosin, and platelet myosin. , 1981, The Journal of biological chemistry.

[31]  J. Spudich,et al.  Control of nonmuscle myosins by phosphorylation. , 1992, Annual review of biochemistry.

[32]  D. Hartshorne,et al.  Calyculin-A increases the level of protein phosphorylation and changes the shape of 3T3 fibroblasts. , 1991, Cell motility and the cytoskeleton.