Myosin light chain kinase stimulates smooth muscle myosin ATPase activity by binding to the myosin heads without phosphorylating the myosin light chain

被引:18
作者
Gao, Y [1 ]
Kawano, K [1 ]
Yoshiyama, S [1 ]
Kawamichi, H [1 ]
Wang, XM [1 ]
Nakamura, A [1 ]
Kohama, K [1 ]
机构
[1] Gunma Univ, Sch Med, Dept Pharmacol, Maebashi, Gumma 3718511, Japan
关键词
myosin light chain kinase; multifunctional regulatory protein; smooth muscle contraction; myosin activation; kinase activity; non-kinase activity; myosin-binding activity; site of action; heavy mero-myosin; subfragment-1;
D O I
10.1016/S0006-291X(03)00690-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Myosin light chain kinase (MLCK) is a multifunctional regulatory protein of smooth muscle contraction [IUBMB Life 51 (2001) 337, for review]. The well-established mode for its regulation is to phosphorylate the 20 kDa myosin light chain (MLC 20) to activate myosin ATPase activity. MLCK exhibits myosin-binding activity in addition to this kinase activity. The myosin-binding activity also stimulates myosin ATPase activity without phosphorylating MLC 20 [Proc. Natl. Acad. Sci. USA 96 (1999) 6666]. We engineered an MLCK fragment containing the myosin-binding domain but devoid of a catalytic domain to explore how myosin is stimulated by this non-kinase pathway. The recombinant fragment thus obtained stimulated myosin ATPase activity by V-max = 5.53 +/- 0.63-fold with K-m = 4.22 +/- 0.58 muM (n = 4). Similar stimulation figures were obtained by measuring the ATPase activity of HMM and S1. Binding of the fragment to both HMM and S1 was also verified, indicating that the fragment exerts stimulation through the myosin heads. Since S I is in an active form regardless of the phosphorylated state of MLC 20, we conclude that the non-kinase stimulation is independent of the phosphorylating mode for activation of myosin. (C) 2003 Elsevier Science (USA). All rights reserved.
引用
收藏
页码:16 / 21
页数:6
相关论文
共 50 条
[41]   Role of myosin light chain and myosin light chain kinase in advanced glycation end product-induced endothelial hyperpermeability in vitro and in vivo [J].
Wu, Fan ;
Guo, Xiaohua ;
Xu, Jing ;
Wang, Weiju ;
Li, Bingling ;
Huang, Qiaobing ;
Su, Lei ;
Xu, Qiulin .
DIABETES & VASCULAR DISEASE RESEARCH, 2016, 13 (02) :137-144
[42]   Myosin light chain kinase: functional domains and structural motifs [J].
Stull, JT ;
Lin, PJ ;
Krueger, JK ;
Trewhella, J ;
Zhi, G .
ACTA PHYSIOLOGICA SCANDINAVICA, 1998, 164 (04) :471-482
[43]   Myosin light chain kinase controls voltage-dependent calcium channels in vascular smooth muscle [J].
Martinsen, A. ;
Schakman, O. ;
Yerna, X. ;
Dessy, C. ;
Morel, N. .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 2014, 466 (07) :1377-1389
[44]   PSEUDOSUBSTRATE SEQUENCE MAY NOT BE CRITICAL FOR AUTOINHIBITION OF SMOOTH-MUSCLE MYOSIN LIGHT-CHAIN KINASE [J].
TANAKA, M ;
IKEBE, R ;
MATSUURA, M ;
IKEBE, M .
EMBO JOURNAL, 1995, 14 (12) :2839-2846
[45]   Myosin light chain kinase controls voltage-dependent calcium channels in vascular smooth muscle [J].
A. Martinsen ;
O. Schakman ;
X. Yerna ;
C. Dessy ;
N. Morel .
Pflügers Archiv - European Journal of Physiology, 2014, 466 :1377-1389
[46]   Localization and activity of myosin light chain kinase isoforms during the cell cycle [J].
Poperechnaya, A ;
Varlamova, O ;
Lin, P ;
Stull, JT ;
Bresnick, AR .
JOURNAL OF CELL BIOLOGY, 2000, 151 (03) :697-707
[47]   Kinetic and motor functions mediated by distinct regions of the regulatory light chain of smooth muscle myosin [J].
Ni, Shaowei ;
Hong, Feng ;
Brewer, Paul D. ;
Ikebe, Mitsuo ;
Onishi, Hirofumi ;
Baker, Jonathan E. ;
Facemyer, Kevin C. ;
Cremo, Christine R. .
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 2009, 1794 (11) :1599-1605
[48]   Effects of pseudophosphorylation mutants on the structural dynamics of smooth muscle myosin regulatory light chain [J].
Espinoza-Fonseca, L. Michel ;
Colson, Brett A. ;
Thomas, David D. .
MOLECULAR BIOSYSTEMS, 2014, 10 (10) :2693-2698
[49]   Effect of vanadate on force and myosin light chain phosphorylation in skinned aortic smooth muscle [J].
Morimoto, S ;
Sato, O ;
Ogawa, Y .
JOURNAL OF BIOCHEMISTRY, 1999, 126 (01) :146-152
[50]   Inhibition of smooth-muscle myosin-light-chain phosphatase by Ruthenium Red [J].
Yamada, A ;
Sato, O ;
Watanabe, M ;
Walsh, MP ;
Ogawa, Y ;
Imaizumi, Y .
BIOCHEMICAL JOURNAL, 2000, 349 :797-804