Thrombin-induced activation of RhoA in platelet shape change

S. L. Bodie, Isobel Ford, Michael Greaves, Graeme Fleming Nixon

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Thrombin-induced activation of RhoA and its involvement in the regulation of myosin II light chain(20) phosphorylation (MLC-P) in alpha -toxin permeabilized platelets was investigated. Permeabilized platelets, expressing normal levels of P-selectin, displayed a Ca2+-dependent increase in shape change and MLC-P. Thrombin activated RhoA as measured by a rhotekin-binding assay within 30 s of stimulation under conditions of constant [Ca2+](i). Under the same conditions and timecourse, thrombin or GTP gammaS induced an increase in MLC-P and platelet shape change which was not dependent on an increase in [Ca2+](i). The thrombin- and GTP gammaS-induced MLC-P in constant [Ca2+](i) was inhibited by the addition of Y27632, a Rho-kinase inhibitor. This study directly demonstrates that thrombin can activate RhoA in platelets in a timecourse compatible with a role in increasing MLC-P and shape change (not involving an increase in [Ca2+](i)). This is also Rho-kinase-dependent. (C) 2001 Academic Press.

Original languageEnglish
Pages (from-to)71-76
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume287
Issue number1
DOIs
Publication statusPublished - 2001

Keywords

  • rho
  • rho-kinase
  • platelet shape change
  • thrombin
  • myosin light chain phosphorylation
  • myosin light-chain
  • binding protein-RHO
  • smooth-muscle
  • kinase
  • phosphorylation
  • phosphatase
  • calcium
  • pathway
  • cytoskeleton
  • secretion

Cite this

Thrombin-induced activation of RhoA in platelet shape change. / Bodie, S. L.; Ford, Isobel; Greaves, Michael; Nixon, Graeme Fleming.

In: Biochemical and Biophysical Research Communications, Vol. 287, No. 1, 2001, p. 71-76.

Research output: Contribution to journalArticle

Bodie, S. L. ; Ford, Isobel ; Greaves, Michael ; Nixon, Graeme Fleming. / Thrombin-induced activation of RhoA in platelet shape change. In: Biochemical and Biophysical Research Communications. 2001 ; Vol. 287, No. 1. pp. 71-76.
@article{6584532f7cf34460a4e376d056eaebee,
title = "Thrombin-induced activation of RhoA in platelet shape change",
abstract = "Thrombin-induced activation of RhoA and its involvement in the regulation of myosin II light chain(20) phosphorylation (MLC-P) in alpha -toxin permeabilized platelets was investigated. Permeabilized platelets, expressing normal levels of P-selectin, displayed a Ca2+-dependent increase in shape change and MLC-P. Thrombin activated RhoA as measured by a rhotekin-binding assay within 30 s of stimulation under conditions of constant [Ca2+](i). Under the same conditions and timecourse, thrombin or GTP gammaS induced an increase in MLC-P and platelet shape change which was not dependent on an increase in [Ca2+](i). The thrombin- and GTP gammaS-induced MLC-P in constant [Ca2+](i) was inhibited by the addition of Y27632, a Rho-kinase inhibitor. This study directly demonstrates that thrombin can activate RhoA in platelets in a timecourse compatible with a role in increasing MLC-P and shape change (not involving an increase in [Ca2+](i)). This is also Rho-kinase-dependent. (C) 2001 Academic Press.",
keywords = "rho, rho-kinase, platelet shape change, thrombin, myosin light chain phosphorylation, myosin light-chain, binding protein-RHO, smooth-muscle, kinase, phosphorylation, phosphatase, calcium, pathway, cytoskeleton, secretion",
author = "Bodie, {S. L.} and Isobel Ford and Michael Greaves and Nixon, {Graeme Fleming}",
year = "2001",
doi = "10.1006/bbrc.2001.5547",
language = "English",
volume = "287",
pages = "71--76",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Thrombin-induced activation of RhoA in platelet shape change

AU - Bodie, S. L.

AU - Ford, Isobel

AU - Greaves, Michael

AU - Nixon, Graeme Fleming

PY - 2001

Y1 - 2001

N2 - Thrombin-induced activation of RhoA and its involvement in the regulation of myosin II light chain(20) phosphorylation (MLC-P) in alpha -toxin permeabilized platelets was investigated. Permeabilized platelets, expressing normal levels of P-selectin, displayed a Ca2+-dependent increase in shape change and MLC-P. Thrombin activated RhoA as measured by a rhotekin-binding assay within 30 s of stimulation under conditions of constant [Ca2+](i). Under the same conditions and timecourse, thrombin or GTP gammaS induced an increase in MLC-P and platelet shape change which was not dependent on an increase in [Ca2+](i). The thrombin- and GTP gammaS-induced MLC-P in constant [Ca2+](i) was inhibited by the addition of Y27632, a Rho-kinase inhibitor. This study directly demonstrates that thrombin can activate RhoA in platelets in a timecourse compatible with a role in increasing MLC-P and shape change (not involving an increase in [Ca2+](i)). This is also Rho-kinase-dependent. (C) 2001 Academic Press.

AB - Thrombin-induced activation of RhoA and its involvement in the regulation of myosin II light chain(20) phosphorylation (MLC-P) in alpha -toxin permeabilized platelets was investigated. Permeabilized platelets, expressing normal levels of P-selectin, displayed a Ca2+-dependent increase in shape change and MLC-P. Thrombin activated RhoA as measured by a rhotekin-binding assay within 30 s of stimulation under conditions of constant [Ca2+](i). Under the same conditions and timecourse, thrombin or GTP gammaS induced an increase in MLC-P and platelet shape change which was not dependent on an increase in [Ca2+](i). The thrombin- and GTP gammaS-induced MLC-P in constant [Ca2+](i) was inhibited by the addition of Y27632, a Rho-kinase inhibitor. This study directly demonstrates that thrombin can activate RhoA in platelets in a timecourse compatible with a role in increasing MLC-P and shape change (not involving an increase in [Ca2+](i)). This is also Rho-kinase-dependent. (C) 2001 Academic Press.

KW - rho

KW - rho-kinase

KW - platelet shape change

KW - thrombin

KW - myosin light chain phosphorylation

KW - myosin light-chain

KW - binding protein-RHO

KW - smooth-muscle

KW - kinase

KW - phosphorylation

KW - phosphatase

KW - calcium

KW - pathway

KW - cytoskeleton

KW - secretion

U2 - 10.1006/bbrc.2001.5547

DO - 10.1006/bbrc.2001.5547

M3 - Article

VL - 287

SP - 71

EP - 76

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -