The effects of the Rho-kinase inhibitor Y-27632 on arachidonic acid-, GTPγS-, and phorbol ester-induced Ca2+-sensitization of smooth muscle

Xiaohong Fu; Ming Cui Gong; Taiping Jia; Avril V. Somlyo; Andrew P. Somlyo

doi:10.1016/S0014-5793(98)01455-0

The effects of the Rho-kinase inhibitor Y-27632 on arachidonic acid-, GTPγS-, and phorbol ester-induced Ca²⁺-sensitization of smooth muscle

Xiaohong Fu, Ming Cui Gong, Taiping Jia, Avril V. Somlyo, Andrew P. Somlyo

Research output: Contribution to journal › Article › peer-review

197 Scopus citations

Abstract

The effects of the Rho-kinase inhibitor, Y-27632 on Ca²⁺-sensitization of force induced by arachidonic acid (AA), phorbol 12,13-dibutyrate (PDBu), GTPγS, and by the stable thromboxane analog, 9,11-dideoxy-9α,11α-methanoepoxy-PGF(2α) (U-46619), were determined in α-toxin-permeabilized smooth muscles. Y-27632 relaxed (up to 99%) Ca²⁺-sensitization by GTPγS (10 μM) and U-46619 (1 μM), but not by PDBu (20 μM), and reduced GTPγS-induced myosin light chain (MLC₂₀) phosphorylation from 28% to 17% (P=0.002). GTPγS-induced force sensitization was inhibited by Y-27632 more potently when the inhibitor was added during the plateau of force than prior to stimulation. In α-toxin-permeabilized smooth muscle, Y-27632 inhibited AA (50 μM)-induced Ca²⁺-sensitization of force (by 66±1.3%) and reduced MLC₂₀ phosphorylation. In contrast, Y-27632 did not relax force Ca²⁺-sensitized by AA in smooth muscle permeabilized with Triton X-100. We conclude that (i) AA induces Ca²⁺-sensitization through dual mechanisms, one mediated by Rho-kinase (or a related kinase), and (ii) Rho-kinase is not required for phorbol ester-induced Ca²⁺-sensitization. Copyright (C) 1998 Federation of European Biochemical Societies.

Original language	English
Pages (from-to)	183-187
Number of pages	5
Journal	FEBS Letters
Volume	440
Issue number	1-2
DOIs	https://doi.org/10.1016/S0014-5793(98)01455-0
State	Published - Nov 27 1998

Bibliographical note

Funding Information:
We thank Dr. S. Murakami, Ms. A. Yoshimura and Mr. Miura of Yoshitomi Pharmaceutical Industries, Ltd. for generous gifts of Y-27632. This work was supported by NIH Grant HL48807 and an American Heart Association National Scientist Development Grant (M.C.G.). We thank Susan Ramos for excellent technical assistance.

Funding

We thank Dr. S. Murakami, Ms. A. Yoshimura and Mr. Miura of Yoshitomi Pharmaceutical Industries, Ltd. for generous gifts of Y-27632. This work was supported by NIH Grant HL48807 and an American Heart Association National Scientist Development Grant (M.C.G.). We thank Susan Ramos for excellent technical assistance.

Funders	Funder number
National Institutes of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)	P01HL048807
American the American Heart Association

Keywords

Kinase C
Phosphatase
Rho
Signal transduction

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

Access to Document

10.1016/S0014-5793(98)01455-0

Cite this

@article{8cf3e72a4a1b41fd9a0b175e1380392d,

title = "The effects of the Rho-kinase inhibitor Y-27632 on arachidonic acid-, GTPγS-, and phorbol ester-induced Ca2+-sensitization of smooth muscle",

abstract = "The effects of the Rho-kinase inhibitor, Y-27632 on Ca2+-sensitization of force induced by arachidonic acid (AA), phorbol 12,13-dibutyrate (PDBu), GTPγS, and by the stable thromboxane analog, 9,11-dideoxy-9α,11α-methanoepoxy-PGF(2α) (U-46619), were determined in α-toxin-permeabilized smooth muscles. Y-27632 relaxed (up to 99\%) Ca2+-sensitization by GTPγS (10 μM) and U-46619 (1 μM), but not by PDBu (20 μM), and reduced GTPγS-induced myosin light chain (MLC20) phosphorylation from 28\% to 17\% (P=0.002). GTPγS-induced force sensitization was inhibited by Y-27632 more potently when the inhibitor was added during the plateau of force than prior to stimulation. In α-toxin-permeabilized smooth muscle, Y-27632 inhibited AA (50 μM)-induced Ca2+-sensitization of force (by 66±1.3\%) and reduced MLC20 phosphorylation. In contrast, Y-27632 did not relax force Ca2+-sensitized by AA in smooth muscle permeabilized with Triton X-100. We conclude that (i) AA induces Ca2+-sensitization through dual mechanisms, one mediated by Rho-kinase (or a related kinase), and (ii) Rho-kinase is not required for phorbol ester-induced Ca2+-sensitization. Copyright (C) 1998 Federation of European Biochemical Societies.",

keywords = "Kinase C, Phosphatase, Rho, Signal transduction",

author = "Xiaohong Fu and Gong, \{Ming Cui\} and Taiping Jia and Somlyo, \{Avril V.\} and Somlyo, \{Andrew P.\}",

note = "Funding Information: We thank Dr. S. Murakami, Ms. A. Yoshimura and Mr. Miura of Yoshitomi Pharmaceutical Industries, Ltd. for generous gifts of Y-27632. This work was supported by NIH Grant HL48807 and an American Heart Association National Scientist Development Grant (M.C.G.). We thank Susan Ramos for excellent technical assistance.",

year = "1998",

month = nov,

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doi = "10.1016/S0014-5793(98)01455-0",

language = "English",

volume = "440",

pages = "183--187",

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TY - JOUR

T1 - The effects of the Rho-kinase inhibitor Y-27632 on arachidonic acid-, GTPγS-, and phorbol ester-induced Ca2+-sensitization of smooth muscle

AU - Fu, Xiaohong

AU - Gong, Ming Cui

AU - Jia, Taiping

AU - Somlyo, Avril V.

AU - Somlyo, Andrew P.

N1 - Funding Information: We thank Dr. S. Murakami, Ms. A. Yoshimura and Mr. Miura of Yoshitomi Pharmaceutical Industries, Ltd. for generous gifts of Y-27632. This work was supported by NIH Grant HL48807 and an American Heart Association National Scientist Development Grant (M.C.G.). We thank Susan Ramos for excellent technical assistance.

PY - 1998/11/27

Y1 - 1998/11/27

N2 - The effects of the Rho-kinase inhibitor, Y-27632 on Ca2+-sensitization of force induced by arachidonic acid (AA), phorbol 12,13-dibutyrate (PDBu), GTPγS, and by the stable thromboxane analog, 9,11-dideoxy-9α,11α-methanoepoxy-PGF(2α) (U-46619), were determined in α-toxin-permeabilized smooth muscles. Y-27632 relaxed (up to 99%) Ca2+-sensitization by GTPγS (10 μM) and U-46619 (1 μM), but not by PDBu (20 μM), and reduced GTPγS-induced myosin light chain (MLC20) phosphorylation from 28% to 17% (P=0.002). GTPγS-induced force sensitization was inhibited by Y-27632 more potently when the inhibitor was added during the plateau of force than prior to stimulation. In α-toxin-permeabilized smooth muscle, Y-27632 inhibited AA (50 μM)-induced Ca2+-sensitization of force (by 66±1.3%) and reduced MLC20 phosphorylation. In contrast, Y-27632 did not relax force Ca2+-sensitized by AA in smooth muscle permeabilized with Triton X-100. We conclude that (i) AA induces Ca2+-sensitization through dual mechanisms, one mediated by Rho-kinase (or a related kinase), and (ii) Rho-kinase is not required for phorbol ester-induced Ca2+-sensitization. Copyright (C) 1998 Federation of European Biochemical Societies.

AB - The effects of the Rho-kinase inhibitor, Y-27632 on Ca2+-sensitization of force induced by arachidonic acid (AA), phorbol 12,13-dibutyrate (PDBu), GTPγS, and by the stable thromboxane analog, 9,11-dideoxy-9α,11α-methanoepoxy-PGF(2α) (U-46619), were determined in α-toxin-permeabilized smooth muscles. Y-27632 relaxed (up to 99%) Ca2+-sensitization by GTPγS (10 μM) and U-46619 (1 μM), but not by PDBu (20 μM), and reduced GTPγS-induced myosin light chain (MLC20) phosphorylation from 28% to 17% (P=0.002). GTPγS-induced force sensitization was inhibited by Y-27632 more potently when the inhibitor was added during the plateau of force than prior to stimulation. In α-toxin-permeabilized smooth muscle, Y-27632 inhibited AA (50 μM)-induced Ca2+-sensitization of force (by 66±1.3%) and reduced MLC20 phosphorylation. In contrast, Y-27632 did not relax force Ca2+-sensitized by AA in smooth muscle permeabilized with Triton X-100. We conclude that (i) AA induces Ca2+-sensitization through dual mechanisms, one mediated by Rho-kinase (or a related kinase), and (ii) Rho-kinase is not required for phorbol ester-induced Ca2+-sensitization. Copyright (C) 1998 Federation of European Biochemical Societies.

KW - Kinase C

KW - Phosphatase

KW - Rho

KW - Signal transduction

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