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Vol. 57, Issue 5, 976-983, May 2000
Department of Pharmacology, Kyoto University, Faculty of Medicine,
Kyoto, Japan (T.I., J.K., K.N., M.M., S.N.); and Drug Discovery
Laboratories (Osaka), WelFide Corporation (Yoshitomi Pharmaceutical
Industries), Hirakata, Japan (M.U., I.T.)
Y-27632
[(+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl)cyclohexanecarboxamide
dihydrochloride] is widely used as a specific inhibitor of the
Rho-associated coiled-coil forming protein serine/threonine kinase
(ROCK) family of protein kinases. This study examined the inhibition
mechanism and profile of actions of Y-27632 and a related compound,
Y-30141
[(+)-(R)-trans- 4-(1-aminoethyl)-N-(1H-pyrrolo[2,3-b]pyridin-4-yl)cyclohexan-ecarboxamide dihydrochloride]. Y-27632 and Y-30141 inhibited the kinase activity of
both ROCK-I and ROCK-II in vitro, and this inhibition was reversed by
ATP in a competitive manner. This suggests that these compounds inhibit
the kinases by binding to the catalytic site. Their affinities for ROCK
kinases as determined by Ki values were at
least 20 to 30 times higher than those for two other Rho effector
kinases, citron kinase and protein kinase PKN.
[3H]Y-30141 was taken up by cells in a temperature- and
time-dependent and saturable manner, and this uptake was competed with
unlabeled Y-27632. No concentrated accumulation was found, suggesting
that the uptake is a carrier-mediated facilitated diffusion. Y-27632 abolished stress fibers in Swiss 3T3 cells at 10 µM, but the
G1-S phase transition of the cell cycle and
cytokinesis were little affected at this concentration. Y-30141 was 10 times more potent than Y-27632 in inhibiting the kinase activity and
stress fiber formation, and it caused significant delay in the
G1-S transition and inhibition of cytokinesis at 10 µM.
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Y. Liu, Y. J. Suzuki, R. M. Day, and B. L. Fanburg Rho Kinase-Induced Nuclear Translocation of ERK1/ERK2 in Smooth Muscle Cell Mitogenesis Caused by Serotonin Circ. Res., September 17, 2004; 95(6): 579 - 586. [Abstract] [Full Text] [PDF]
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A. M. Avalos, W. T. Arthur, P. Schneider, A. F. G. Quest, K. Burridge, and L. Leyton Aggregation of Integrins and RhoA Activation Are Required for Thy-1-induced Morphological Changes in Astrocytes J. Biol. Chem., September 10, 2004; 279(37): 39139 - 39145. [Abstract] [Full Text] [PDF]
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 F. D Russell and P. Molenaar Investigation of signaling pathways that mediate the inotropic effect of urotensin-II in human heart Cardiovasc Res, September 1, 2004; 63(4): 673 - 681. [Abstract] [Full Text] [PDF]
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K. Budzyn, P. D. Marley, and C. G. Sobey Chronic mevastatin modulates receptor-dependent vascular contraction in eNOS-deficient mice Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2004; 287(2): R342 - R348. [Abstract] [Full Text] [PDF]
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 T. Kamai, T. Yamanishi, H. Shirataki, K. Takagi, H. Asami, Y. Ito, and K.-I. Yoshida Overexpression of RhoA, Rac1, and Cdc42 GTPases Is Associated with Progression in Testicular Cancer Clin. Cancer Res., July 15, 2004; 10(14): 4799 - 4805. [Abstract] [Full Text] [PDF]
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 S. A. K. Harvey, S. C. Anderson, and N. SundarRaj Downstream Effects of ROCK Signaling in Cultured Human Corneal Stromal Cells: Microarray Analysis of Gene Expression Invest. Ophthalmol. Vis. Sci., July 1, 2004; 45(7): 2168 - 2176. [Abstract] [Full Text] [PDF]
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P. S. Mettu, P.-F. Deng, U. K. Misra, G. Gawdi, D. L. Epstein, and P. V. Rao Role of Lysophospholipid Growth Factors in the Modulation of Aqueous Humor Outflow Facility Invest. Ophthalmol. Vis. Sci., July 1, 2004; 45(7): 2263 - 2271. [Abstract] [Full Text] [PDF]
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N. Teusch, E. Lombardo, J. Eddleston, and U. G. Knaus The Low Molecular Weight GTPase RhoA and Atypical Protein Kinase C{zeta} Are Required for TLR2-Mediated Gene Transcription J. Immunol., July 1, 2004; 173(1): 507 - 514. [Abstract] [Full Text] [PDF]
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F. Causeret, M. Hidalgo-Sanchez, P. Fort, S. Backer, M.-R. Popoff, C. Gauthier-Rouviere, and E. Bloch-Gallego Distinct roles of Rac1/Cdc42 and Rho/Rock for axon outgrowth and nucleokinesis of precerebellar neurons toward netrin 1 Development, June 15, 2004; 131(12): 2841 - 2852. [Abstract] [Full Text] [PDF]
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Z. A. Pfeiffer, M. Aga, U. Prabhu, J. J. Watters, D. J. Hall, and P. J. Bertics The nucleotide receptor P2X7 mediates actin reorganization and membrane blebbing in RAW 264.7 macrophages via p38 MAP kinase and Rho J. Leukoc. Biol., June 1, 2004; 75(6): 1173 - 1182. [Abstract] [Full Text] [PDF]
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S.R. Bailey, A.H. Eid, S. Mitra, S. Flavahan, and N.A. Flavahan Rho Kinase Mediates Cold-Induced Constriction of Cutaneous Arteries: Role of {alpha}2C-Adrenoceptor Translocation Circ. Res., May 28, 2004; 94(10): 1367 - 1374. [Abstract] [Full Text] [PDF]
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 C. V. Melendez-Vasquez, S. Einheber, and J. L. Salzer Rho Kinase Regulates Schwann Cell Myelination and Formation of Associated Axonal Domains J. Neurosci., April 21, 2004; 24(16): 3953 - 3963. [Abstract] [Full Text] [PDF]
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H. Honing, T. K. van den Berg, S. M. A. van der Pol, C. D. Dijkstra, R. A. van der Kammen, J. G. Collard, and H. E. de Vries RhoA activation promotes transendothelial migration of monocytes via ROCK J. Leukoc. Biol., March 1, 2004; 75(3): 523 - 528. [Abstract] [Full Text] [PDF]
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M. Moore, B. A. Marroquin, W. Gugliotta, R. Tse, and S. R. White Rho Kinase Inhibition Initiates Apoptosis in Human Airway Epithelial Cells Am. J. Respir. Cell Mol. Biol., March 1, 2004; 30(3): 379 - 387. [Abstract] [Full Text] [PDF]
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M. V. Hoang, M. C. Whelan, and D. R. Senger Rho activity critically and selectively regulates endothelial cell organization during angiogenesis PNAS, February 17, 2004; 101(7): 1874 - 1879. [Abstract] [Full Text] [PDF]
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D. A. Emmert, J. A. Fee, Z. M. Goeckeler, J. M. Grojean, T. Wakatsuki, E. L. Elson, B. P. Herring, P. J. Gallagher, and R. B. Wysolmerski Rho-kinase-mediated Ca2+-independent contraction in rat embryo fibroblasts Am J Physiol Cell Physiol, January 1, 2004; 286(1): C8 - C21. [Abstract] [Full Text] [PDF]
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H. Setiadi and R. P. McEver Signal-dependent distribution of cell surface P-selectin in clathrin-coated pits affects leukocyte rolling under flow J. Cell Biol., December 22, 2003; 163(6): 1385 - 1395. [Abstract] [Full Text] [PDF]
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A.-C. Tosello-Trampont, K. Nakada-Tsukui, and K. S. Ravichandran Engulfment of Apoptotic Cells Is Negatively Regulated by Rho-mediated Signaling J. Biol. Chem., December 12, 2003; 278(50): 49911 - 49919. [Abstract] [Full Text] [PDF]
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 F. E. Nwariaku, P. Rothenbach, Z. Liu, X. Zhu, R. H. Turnage, and L. S. Terada Rho inhibition decreases TNF-induced endothelial MAPK activation and monolayer permeability J Appl Physiol, November 1, 2003; 95(5): 1889 - 1895. [Abstract] [Full Text] [PDF]
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H. Ikeda, K. Nagashima, M. Yanase, T. Tomiya, M. Arai, Y. Inoue, K. Tejima, T. Nishikawa, M. Omata, S. Kimura, et al. Involvement of Rho/Rho kinase pathway in regulation of apoptosis in rat hepatic stellate cells Am J Physiol Gastrointest Liver Physiol, November 1, 2003; 285(5): G880 - G886. [Abstract] [Full Text] [PDF]
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W. Bing, S. Chang, J. A. Hypolite, M. E. DiSanto, S. A. Zderic, L. Rolf, A. J. Wein, and S. Chacko Obstruction-induced changes in urinary bladder smooth muscle contractility: a role for Rho kinase Am J Physiol Renal Physiol, November 1, 2003; 285(5): F990 - F997. [Abstract] [Full Text] [PDF]
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Z. Mallat, A. Gojova, V. Sauzeau, V. Brun, J.-S. Silvestre, B. Esposito, R. Merval, H. Groux, G. Loirand, and A. Tedgui Rho-Associated Protein Kinase Contributes to Early Atherosclerotic Lesion Formation in Mice Circ. Res., October 31, 2003; 93(9): 884 - 888. [Abstract] [Full Text] [PDF]
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 A. P. SOMLYO and A. V. SOMLYO Ca2+ Sensitivity of Smooth Muscle and Nonmuscle Myosin II: Modulated by G Proteins, Kinases, and Myosin Phosphatase Physiol Rev, October 1, 2003; 83(4): 1325 - 1358. [Abstract] [Full Text] [PDF]
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J. S. Da Silva, M. Medina, C. Zuliani, A. Di Nardo, W. Witke, and C. G. Dotti RhoA/ROCK regulation of neuritogenesis via profilin IIa-mediated control of actin stability J. Cell Biol., September 29, 2003; 162(7): 1267 - 1279. [Abstract] [Full Text] [PDF]
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M. Zhang, G. M. Kalinec, R. Urrutia, D. D. Billadeau, and F. Kalinec ROCK-dependent and ROCK-independent Control of Cochlear Outer Hair Cell Electromotility J. Biol. Chem., September 12, 2003; 278(37): 35644 - 35650. [Abstract] [Full Text] [PDF]
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C. L. Gatto, B. J. Walker, and S. Lambert Local ERM activation and dynamic growth cones at Schwann cell tips implicated in efficient formation of nodes of Ranvier J. Cell Biol., August 4, 2003; 162(3): 489 - 498. [Abstract] [Full Text] [PDF]
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M. A. Chellaiah, R. S. Biswas, S. R. Rittling, D. T. Denhardt, and K. A. Hruska Rho-dependent Rho Kinase Activation Increases CD44 Surface Expression and Bone Resorption in Osteoclasts J. Biol. Chem., August 1, 2003; 278(31): 29086 - 29097. [Abstract] [Full Text] [PDF]
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P.-L. Tharaux, R. C. Bukoski, P. N. Rocha, S. D. Crowley, P. Ruiz, C. Nataraj, D. N. Howell, K. Kaibuchi, R. F. Spurney, and T. M. Coffman Rho Kinase Promotes Alloimmune Responses by Regulating the Proliferation and Structure of T Cells J. Immunol., July 1, 2003; 171(1): 96 - 105. [Abstract] [Full Text] [PDF]
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