title: Protein kinase network in the regulation of phosphorylation and dephosphorylation of smooth muscle myosin light chain.
abstract: The contraction of smooth muscle is regulated primarily by intracellular Ca2+ signal. It is well established that the elevation of the cytosolic Ca2+ level activates myosin light chain kinase, which phosphorylates 20 kDa regulatory myosin light chain and activates myosin ATPase. The simultaneous measurement of cytosolic Ca2+ concentration and force development revealed that the alteration of the Ca2+-sensitivity of the contractile apparatus as well as the Ca2+ signal plays a critical role in the regulation of smooth muscle contraction. The fluctuation of an extent of myosin phosphorylation for a given change in Ca2+ concentration is considered to contribute to the major mechanisms regulating the Ca2+-sensitivity. The level of myosin phosphorylation is determined by the balance between phosphorylation and dephosphorylation. The phosphorylation level for a given Ca2+ elevation is increased either by Ca2+-independent activation of phosphorylation process or inhibition of dephosphorylation. In the last decade, the isolation and cloning of myosin phosphatase facilitated the understanding of regulatory mechanism of dephosphorylation process at the molecular level. The inhibition of myosin phosphatase can be achieved by (1) alteration of hetrotrimeric structure, (2) phosphorylation of 110 kDa regulatory subunit MYPT1 at the specific site and (3) inhibitory protein CPI-17 upon its phosphorylation. Rho-kinase was first identified to phosphorylate MYPT1, and later many kinases were found to phosphorylate MYPT1 and inhibit dephosphorylation of myosin. Similarly, the phosphorylation of CPI-17 can be catalysed by multiple kinases. Moreover, the myosin light chain can be phosphorylated by not only authentic myosin light chain kinase in a Ca2+-dependent manner but also by multiple kinases in a Ca2+-independent manner, thus adding a novel mechanism to the regulation of the Ca2+-sensitivity by regulating the phosphorylation process. It is now clarified that the protein kinase network is involved in the regulation of myosin phosphorylation and dephosphorylation. However, the physiological role of each component remains to be determined. One approach to accomplish this purpose is to investigate the effects of the dominant negative mutants of the signalling molecule on the smooth muscle contraction. In this regards, a protein transduction technique utilizing the cell-penetrating peptides would provide a useful tool. In the preliminary study, we succeeded in introducing a fragment of MYPT1 into the arterial strips, and found enhancement of contraction.
authors.affiliation: Division of Molecular Cardiology, Research Institute ofAngiocardiology, Graduate School of Medical Sciences, Kyushu University; Maidashi, Higashi-ku, Fukuoka, Japan. khirano@molcar.med.kyushu-u.ac.jp
authors.forename: Katusya
authors.initials: K
authors.lastname: Hirano
authors.forename: Dmitry N
authors.initials: DN
authors.lastname: Derkach
authors.forename: Mayumi
authors.initials: M
authors.lastname: Hirano
authors.forename: Junji
authors.initials: J
authors.lastname: Nishimura
authors.forename: Hideo
authors.initials: H
authors.lastname: Kanaide
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D019281:Dimerization; D005799:Genes, Dominant; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008954:Models, Biological; D009119:Muscle Contraction; D009124:Muscle Proteins / Q000096:biosynthesis; D009130:Muscle, Smooth / Q000378:metabolism*; D009154:Mutation; D018994:Myosin Light Chains / Q000378:metabolism*; D010749:Phosphoprotein Phosphatases; D010750:Phosphoproteins / Q000096:biosynthesis; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors; D010766:Phosphorylation; D011494:Protein Kinases / Q000502:physiology*; D015398:Signal Transduction; D013997:Time Factors
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D009124:Muscle Proteins; D018994:Myosin Light Chains; C107507:PPP1R14A protein, human; D010750:Phosphoproteins; D011494:Protein Kinases; D010749:Phosphoprotein Phosphatases; D010744:Phosphoric Monoester Hydrolases; D000251:Adenosine Triphosphatases; D002118:Calcium