Detection of myosin light chain phosphorylation-A cell-based assay for screening Rho-kinase inhibitors

Biochemical and Biophysical Research Communications

Volumn 374, Issue 2, 2008, Pages 356-360

  Schröter, Thomas ^a   Griffin, Evelyn ^a   Weiser, Amiee ^a   Feng, Yangbo ^a   LoGrasso, Philip ^a  

^a SCRIPPS RESEARCH INSTITUTE   (United States)

Author keywords

Cell based assay; MLC; Myosin light chain; Phosphorylation; Rho kinase; ROCK; Ser19; Thr18

Indexed keywords

4 (1 AMINOETHYL) N (4 PYRIDYL)CYCLOHEXANECARBOXAMIDE; FASUDIL; H 1152P; MYOSIN LIGHT CHAIN; RHO KINASE INHIBITOR;

ANIMAL CELL; ARTICLE; CELL ASSAY; CELL LINE; CONTROLLED STUDY; CORRELATION ANALYSIS; DRUG SCREENING; ELECTROPHORESIS; ENZYME INHIBITION; IC 50; IMAGING; MEDICAL TECHNOLOGY; NONHUMAN; PHENOTYPE; PRIORITY JOURNAL; PROTEIN PHOSPHORYLATION; QUANTITATIVE ANALYSIS; RAT; SIGNAL TRANSDUCTION;

AMIDES; ANIMALS; CELL LINE; DRUG EVALUATION, PRECLINICAL; MYOSIN LIGHT CHAINS; PHOSPHOPROTEINS; PHOSPHORYLATION; PROTEIN KINASE INHIBITORS; PYRIDINES; RHO-ASSOCIATED KINASES;

EID: 48549099363     PISSN: 0006291X     EISSN: 10902104     Source Type: Journal    
DOI: 10.1016/j.bbrc.2008.07.028     Document Type: Article

References (25)

1. Ikebe M., Koretz J., and Hartshorne D.J. Effects of phosphorylation of light chain residues threonine 18 and serine 19 on the properties and conformation of smooth muscle myosin. J. Biol. Chem. 263 (1988) 6432-6437
2. Iwasaki T., Murata-Hori M., Ishitobi S., and Hosoya H. Diphosphorylated MRLC is required for organization of stress fibers in interphase cells and the contractile ring in dividing cells. Cell Struct. Funct. 26 (2001) 677-683
3. Amano M., Chihara K., Nakamura N., Fukata Y., Yano T., Shibata M., Ikebe M., and Kaibuchi K. Myosin II activation promotes neurite retraction during the action of Rho and Rho-kinase. Genes Cells 3 (1998) 177-188
4. Ueda K., Murata-Hori M., Tatsuka M., and Hosoya H. Rho-kinase contributes to diphosphorylation of myosin II regulatory light chain in nonmuscle cells. Oncogene 21 (2002) 5852-5860
5. Kimura K., Ito M., Amano M., Chihara K., Fukata Y., Nakafuku M., Yamamori B., Feng J., Nakano T., Okawa K., Iwamatsu A., and Kaibuchi K. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase). Science 273 (1996) 245-248
6. Komatsu S., and Ikebe M. ZIP kinase is responsible for the phosphorylation of myosin II and necessary for cell motility in mammalian fibroblasts. J. Cell Biol. 165 (2004) 243-254
7. Yoneda A., Multhaupt H.A., and Couchman J.R. The Rho kinases I and II regulate different aspects of myosin II activity. J. Cell Biol. 170 (2005) 443-453
8. Birukova A.A., Smurova K., Birukov K.G., Kaibuchi K., Garcia J.G.N., and Verin A.D. Role of Rho GTPases in thrombin-induced lung vascular endothelial cells barrier dysfunction. Microvasc. Res. 67 (2004) 64-77
9. Buyukafsar K., and Un I. Effects of the Rho-kinase inhibitors, Y-27632 and fasudil, on the corpus cavernosum from diabetic mice. Eur. J. Pharmacol. 472 (2003) 235-238
10. Price R.D., Oe T., Yamaji T., and Matsuoka N. A simple, flexible, nonfluorescent system for the automated screening of neurite outgrowth. J. Biomol. Screen. (2005)
11. Wylie S.R., and Chantler P.D. Myosin IIA drives neurite retraction. Mol. Biol. Cell 14 (2003) 4654-4666
12. Nagaoka T., Fagan K.A., Gebb S.A., Morris K.G., Suzuki T., Shimokawa H., McMurtry I.F., and Oka M. Inhaled Rho kinase inhibitors are potent and selective vasodilators in rat pulmonary hypertension. Am. J. Respir. Crit. Care Med. 171 (2005) 494-499
13. Uehata M., Ishizaki T., Satoh H., Ono T., Kawahara T., Morishita T., Tamakawa H., Yamagami K., Inui J., Maekawa M., and Narumiya S. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature 389 (1997) 990-994
14. Takahara A., Sugiyama A., Satoh Y., Yoneyama M., and Hashimoto K. Cardiovascular effects of Y-27632, a selective Rho-associated kinase inhibitor, assessed in the halothane-anesthetized canine model. Eur. J. Pharmacol. 460 (2003) 51-57
15. Honjo M., Tanihara H., Inatani M., Kido N., Sawamura T., Yue B.Y.J.T., Narumiya S., and Honda Y. Effects of Rho-associated protein kinase inhibitor Y-27632 on intraocular pressure and outflow facility. Invest. Ophthalmol. Vis. Sci. 42 (2001) 137-144
16. Ying H., Biroc S.L., Li W.-w., Alicke B., Xuan J.-A., Pagila R., Ohashi Y., Okada T., Kamata Y., and Dinter H. The Rho kinase inhibitor fasudil inhibits tumor progression in human and rat tumor models. Mol. Cancer Ther. 5 (2006) 2158-2164
17. Sakurada K., Seto M., and Sasaki Y. Dynamics of myosin light chain phosphorylation at Ser19 and Thr18/Ser19 in smooth muscle cells in culture. Am. J. Physiol. Cell Physiol. 274 (1998) C1563-C1572
18. Watanabe T., Hosoya H., and Yonemura S. Regulation of myosin II dynamics by phosphorylation and dephosphorylation of its light chain in epithelial cells. Mol. Biol. Cell 18 (2007) 605-616
19. Katoh H., Aoki J., Yamaguchi Y., Kitano Y., Ichikawa A., and Negishi M. Constitutively active Galpha12, Galpha13, and Galphaq induce Rho-dependent neurite retraction through different signaling pathways. J. Biol. Chem. 273 (1998) 28700-28707
20. Feng Y., Cameron M.D., Frackowiak B., Griffin E., Lin L., Ruiz C., Schroter T., and LoGrasso P. Structure-activity relationships, and drug metabolism and pharmacokinetic properties for indazole piperazine and indazole piperidine inhibitors of ROCK-II. Bioorg. Med. Chem. Lett. 17 (2007) 2355-2360
21. Yamada A., Hirose K., Hashimoto A., and Iino M. Real-time imaging of myosin II regulatory light-chain phosphorylation using a new protein biosensor. Biochem. J. 385 (2005) 589-594
22. Bhadriraju K., Elliott J.T., Nguyen M., and Plant A.L. Quantifying myosin light chain phosphorylation in single adherent cells with automated fluorescence microscopy. BMC Cell Biol. 8 (2007) 43
23. Birukova A.A., Smurova K., Birukov K.G., Usatyuk P., Liu F., Kaibuchi K., Ricks-Cord A., Natarajan V., Alieva I., Garcia J.G., and Verin A.D. Microtubule disassembly induces cytoskeletal remodeling and lung vascular barrier dysfunction: role of Rho-dependent mechanisms. J. Cell. Physiol. 201 (2004) 55-70
24. Nakayama M., Amano M., Katsumi A., Kaneko T., Kawabata S., Takefuji M., and Kaibuchi K. Rho-kinase and myosin II activities are required for cell type and environment specific migration. Genes Cells 10 (2005) 107-117
25. Nakagawa O., Fujisawa K., Ishizaki T., Saito Y., Nakao K., and Narumiya S. ROCK-I and ROCK-II, two isoforms of Rho-associated coiled-coil forming protein serine/threonine kinase in mice. FEBS Lett. 392 (1996) 189-193