Regulation of endothelial cell myosin light chain phosphorylation and permeability by vanadate

Lydia I. Gilbert-McClain, Alexander Dmitriyevich Verin, Shu Shi, Robert P. Irwin, Joe G.N. Garcia

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The involvement of tyrosine protein phosphorylation in the regulation of endothelial cell (EC) contraction and barrier function is poorly understood. We have previously shown that myosin light chain (MLC) phosphorylation catalyzed by a novel 214 kDa EC myosin light chain kinase (MLCK) isoform is a key event in EC contraction and barrier dysfunction [Garcia et al. (1995): J Cell Physiol 163:510-522; Garcia et al. (1997): Am J Respir Cell Mol Biol 16:487-491]. In this study, we tested the hypothesis that tyrosine phosphatases participate in the regulation of EC contraction and barrier function via modulation of MLCK activity. The tyrosine phosphatase inhibitor, sodium orthovanadate (vanadate), significantly decreased electrical resistance across bovine EC monolayers and increased albumin permeability consistent with EC barrier impairment. Vanadate significantly increased EC MLC phosphorylation in a time-dependent manner (maximal increase observed at 10 min) and augmented both the MLC phosphorylation and permeability responses produced by thrombin, an agonist which rapidly increases tyrosine kinase activities. The vanadate-mediated increase in MLC phosphorylation was not associated with alterations in either phosphorylase A Ser/Thr phosphatase activities or in cytosolic [Ca2+] but was strongly associated with significant increases in EC MLCK phosphotyrosine content. These data suggest that tyrosine phosphatase activities may participate in EC contractile and barrier responses via the regulation of the tyrosine phosphorylation status of EC MLCK.

Original languageEnglish (US)
Pages (from-to)141-155
Number of pages15
JournalJournal of cellular biochemistry
Volume70
Issue number1
DOIs
StatePublished - Jul 1 1998
Externally publishedYes

Fingerprint

Myosin Light Chains
Phosphorylation
Vanadates
Endothelial cells
Permeability
Endothelial Cells
Myosin-Light-Chain Kinase
Tyrosine
Phosphoric Monoester Hydrolases
Phosphorylases
Phosphotyrosine
Acoustic impedance
Electric Impedance
Thrombin
Protein-Tyrosine Kinases
Albumins
Monolayers
Protein Isoforms
Sodium
Modulation

Keywords

  • Endothelial cell
  • MLCK
  • Permeability
  • Tyrosine phosphatase
  • Vanadate

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Regulation of endothelial cell myosin light chain phosphorylation and permeability by vanadate. / Gilbert-McClain, Lydia I.; Verin, Alexander Dmitriyevich; Shi, Shu; Irwin, Robert P.; Garcia, Joe G.N.

In: Journal of cellular biochemistry, Vol. 70, No. 1, 01.07.1998, p. 141-155.

Research output: Contribution to journalArticle

Gilbert-McClain, Lydia I. ; Verin, Alexander Dmitriyevich ; Shi, Shu ; Irwin, Robert P. ; Garcia, Joe G.N. / Regulation of endothelial cell myosin light chain phosphorylation and permeability by vanadate. In: Journal of cellular biochemistry. 1998 ; Vol. 70, No. 1. pp. 141-155.
@article{a6b221b356f340f5a51b16732985d7f4,
title = "Regulation of endothelial cell myosin light chain phosphorylation and permeability by vanadate",
abstract = "The involvement of tyrosine protein phosphorylation in the regulation of endothelial cell (EC) contraction and barrier function is poorly understood. We have previously shown that myosin light chain (MLC) phosphorylation catalyzed by a novel 214 kDa EC myosin light chain kinase (MLCK) isoform is a key event in EC contraction and barrier dysfunction [Garcia et al. (1995): J Cell Physiol 163:510-522; Garcia et al. (1997): Am J Respir Cell Mol Biol 16:487-491]. In this study, we tested the hypothesis that tyrosine phosphatases participate in the regulation of EC contraction and barrier function via modulation of MLCK activity. The tyrosine phosphatase inhibitor, sodium orthovanadate (vanadate), significantly decreased electrical resistance across bovine EC monolayers and increased albumin permeability consistent with EC barrier impairment. Vanadate significantly increased EC MLC phosphorylation in a time-dependent manner (maximal increase observed at 10 min) and augmented both the MLC phosphorylation and permeability responses produced by thrombin, an agonist which rapidly increases tyrosine kinase activities. The vanadate-mediated increase in MLC phosphorylation was not associated with alterations in either phosphorylase A Ser/Thr phosphatase activities or in cytosolic [Ca2+] but was strongly associated with significant increases in EC MLCK phosphotyrosine content. These data suggest that tyrosine phosphatase activities may participate in EC contractile and barrier responses via the regulation of the tyrosine phosphorylation status of EC MLCK.",
keywords = "Endothelial cell, MLCK, Permeability, Tyrosine phosphatase, Vanadate",
author = "Gilbert-McClain, {Lydia I.} and Verin, {Alexander Dmitriyevich} and Shu Shi and Irwin, {Robert P.} and Garcia, {Joe G.N.}",
year = "1998",
month = "7",
day = "1",
doi = "10.1002/(SICI)1097-4644(19980701)70:1<141::AID-JCB14>3.0.CO;2-S",
language = "English (US)",
volume = "70",
pages = "141--155",
journal = "Journal of Cellular Biochemistry",
issn = "0730-2312",
publisher = "Wiley-Liss Inc.",
number = "1",

}

TY - JOUR

T1 - Regulation of endothelial cell myosin light chain phosphorylation and permeability by vanadate

AU - Gilbert-McClain, Lydia I.

AU - Verin, Alexander Dmitriyevich

AU - Shi, Shu

AU - Irwin, Robert P.

AU - Garcia, Joe G.N.

PY - 1998/7/1

Y1 - 1998/7/1

N2 - The involvement of tyrosine protein phosphorylation in the regulation of endothelial cell (EC) contraction and barrier function is poorly understood. We have previously shown that myosin light chain (MLC) phosphorylation catalyzed by a novel 214 kDa EC myosin light chain kinase (MLCK) isoform is a key event in EC contraction and barrier dysfunction [Garcia et al. (1995): J Cell Physiol 163:510-522; Garcia et al. (1997): Am J Respir Cell Mol Biol 16:487-491]. In this study, we tested the hypothesis that tyrosine phosphatases participate in the regulation of EC contraction and barrier function via modulation of MLCK activity. The tyrosine phosphatase inhibitor, sodium orthovanadate (vanadate), significantly decreased electrical resistance across bovine EC monolayers and increased albumin permeability consistent with EC barrier impairment. Vanadate significantly increased EC MLC phosphorylation in a time-dependent manner (maximal increase observed at 10 min) and augmented both the MLC phosphorylation and permeability responses produced by thrombin, an agonist which rapidly increases tyrosine kinase activities. The vanadate-mediated increase in MLC phosphorylation was not associated with alterations in either phosphorylase A Ser/Thr phosphatase activities or in cytosolic [Ca2+] but was strongly associated with significant increases in EC MLCK phosphotyrosine content. These data suggest that tyrosine phosphatase activities may participate in EC contractile and barrier responses via the regulation of the tyrosine phosphorylation status of EC MLCK.

AB - The involvement of tyrosine protein phosphorylation in the regulation of endothelial cell (EC) contraction and barrier function is poorly understood. We have previously shown that myosin light chain (MLC) phosphorylation catalyzed by a novel 214 kDa EC myosin light chain kinase (MLCK) isoform is a key event in EC contraction and barrier dysfunction [Garcia et al. (1995): J Cell Physiol 163:510-522; Garcia et al. (1997): Am J Respir Cell Mol Biol 16:487-491]. In this study, we tested the hypothesis that tyrosine phosphatases participate in the regulation of EC contraction and barrier function via modulation of MLCK activity. The tyrosine phosphatase inhibitor, sodium orthovanadate (vanadate), significantly decreased electrical resistance across bovine EC monolayers and increased albumin permeability consistent with EC barrier impairment. Vanadate significantly increased EC MLC phosphorylation in a time-dependent manner (maximal increase observed at 10 min) and augmented both the MLC phosphorylation and permeability responses produced by thrombin, an agonist which rapidly increases tyrosine kinase activities. The vanadate-mediated increase in MLC phosphorylation was not associated with alterations in either phosphorylase A Ser/Thr phosphatase activities or in cytosolic [Ca2+] but was strongly associated with significant increases in EC MLCK phosphotyrosine content. These data suggest that tyrosine phosphatase activities may participate in EC contractile and barrier responses via the regulation of the tyrosine phosphorylation status of EC MLCK.

KW - Endothelial cell

KW - MLCK

KW - Permeability

KW - Tyrosine phosphatase

KW - Vanadate

UR - http://www.scopus.com/inward/record.url?scp=0032127329&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032127329&partnerID=8YFLogxK

U2 - 10.1002/(SICI)1097-4644(19980701)70:1<141::AID-JCB14>3.0.CO;2-S

DO - 10.1002/(SICI)1097-4644(19980701)70:1<141::AID-JCB14>3.0.CO;2-S

M3 - Article

C2 - 9632115

AN - SCOPUS:0032127329

VL - 70

SP - 141

EP - 155

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 1

ER -