Open AccessJournal Article
Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases.
Toshimasa Ishizaki,Masayoshi Uehata,Ichiro Tamechika,Jeongsin Keel,Kimiko Nonomura,Midori Maekawa,Shuh Narumiya +6 more
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TLDR
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, suggesting that these compounds inhibit the kinases by binding to the catalytic site.Abstract:
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 K(i) values were at least 20 to 30 times higher than those for two other Rho effector kinases, citron kinase and protein kinase PKN. [(3)H]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 microM, but the G(1)-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 G(1)-S transition and inhibition of cytokinesis at 10 microM.read more
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Cell shape, cytoskeletal tension, and rhoa regulate stem cell lineage commitment
TL;DR: It is demonstrated that cell shape regulates commitment of human mesenchymal stem cells to adipocyte or osteoblast fate and mechanical cues experienced in developmental and adult contexts, embodied by cell shape, cytoskeletal tension, and RhoA signaling, are integral to the commitment of stem cell fate.
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A ROCK inhibitor permits survival of dissociated human embryonic stem cells
Kiichi Watanabe,Morio Ueno,Daisuke Kamiya,Ayaka Nishiyama,Michiru Matsumura,Takafumi Wataya,Jun Takahashi,Satomi Nishikawa,Shin-Ichi Nishikawa,Keiko Muguruma,Yoshiki Sasai +10 more
TL;DR: Application of a selective Rho-associated kinase (ROCK) inhibitor, Y-27632, to hES cells markedly diminishes dissociation-induced apoptosis, increases cloning efficiency and facilitates subcloning after gene transfer, and enables SFEB-cultured hES Cells to survive and differentiate into Bf1+ cortical and basal telencephalic progenitors.
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Ca2+ Sensitivity of Smooth Muscle and Nonmuscle Myosin II: Modulated by G Proteins, Kinases, and Myosin Phosphatase
Andrew P. Somlyo,Avril V. Somlyo +1 more
TL;DR: It is suggested that the RhoA/ROK pathway is constitutively active in a number of organs under physiological conditions; its aberrations play major roles in several disease states, particularly impacting on Ca2+ sensitization of smooth muscle in hypertension and possibly asthma and on cancer neoangiogenesis and cancer progression.
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ROCKs: multifunctional kinases in cell behaviour
TL;DR: Recent experiments have defined new functions of ROCKs in cells, including centrosome positioning and cell-size regulation, which might contribute to various physiological and pathological states.
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Caspase-3-mediated cleavage of ROCK I induces MLC phosphorylation and apoptotic membrane blebbing
Michael Sebbagh,Claire Renvoizé,Jocelyne Hamelin,Nicole Riche,Jacques Bertoglio,Jacqueline Bréard +5 more
TL;DR: Activation of ROCK I by caspase-3 seems to be responsible for bleb formation in apoptotic cells.
References
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Journal ArticleDOI
Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension
Masayoshi Uehata,Toshimasa Ishizaki,Hiroyuki Satoh,Takashi Ono,Toshio Kawahara,Tamami Morishita,Hiroki Tamakawa,Keiji Yamagami,Jun Inui,Midori Maekawa,Shuh Narumiya +10 more
TL;DR: Pyridine derivative Y-27632 consistently suppresses Rho-induced, p160ROCK-mediated formation of stress fibres in cultured cells and dramatically corrects hypertension in several hypertensive rat models, suggesting that compounds that inhibit this process might be useful therapeutically.
Journal ArticleDOI
Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase)
Kazushi Kimura,Masaaki Ito,Mutsuki Amano,Kazuyasu Chihara,Yuko Fukata,Masato Nakafuku,Bunpei Yamamori,Jianhua Feng,Takeshi Nakano,Katsuya Okawa,Akihiro Iwamatsu,Kozo Kaibuchi +11 more
TL;DR: Rho appears to inhibit myosin phosphatase through the action of Rho-kinase, which is activated by GTP·RhoA, phosphorylation of MBS and MLC in NIH 3T3 cells.
Journal ArticleDOI
Physiological concentrations of purines and pyrimidines.
TL;DR: Consideration of experiments on the intracellular compartmentation of nucleotides shows support for this process between the cytoplasm and mitochondria, but not between the cytoskeleton and the nucleus.
Journal ArticleDOI
Rho GTPases Control Polarity, Protrusion, and Adhesion during Cell Movement
Catherine D. Nobes,Alan Hall +1 more
TL;DR: It is concluded that the signal transduction pathways controlled by the four small GTPases, Rho, Rac, Cdc42, and Ras, cooperate to promote cell movement.
Journal ArticleDOI
Structures of the tyrosine kinase domain of fibroblast growth factor receptor in complex with inhibitors.
Moosa Mohammadi,Gerald McMahon,Li Sun,Cho Tang,Peter Hirth,Brian K. Yeh,Stevan R. Hubbard,Joseph Schlessinger +7 more
TL;DR: A new class of protein tyrosine kinase inhibitors was identified that is based on an oxindole core (indolinones) and two compounds from this class inhibited the kinase activity of fibroblast growth factor receptor 1 (FGFR1) and showed differential specificity toward other receptor tyrosin kinases.