Tyrosine kinase activity, cytoskeletal organization, and motility in human vascular endothelial cells

Lewis H. Romer, Natalie McLean, Christopher E. Turner, Keith Burridge

Research output: Contribution to journalArticlepeer-review

Abstract

Tyrosine phosphorylation of cytoskeletal proteins occurs during integrin- mediated cell adhesion to extracellular matrix proteins. We have investigated the role of tyrosine phosphorylation in the migration and initial spreading of human umbilical vein endothelial cells (HUVEC). Elevated phosphotyrosine concentrations were noted in the focal adhesions of HUVEC migrating into wounds. Anti-phosphotyrosine Western blots of extracts of wounded HUVEC monolayers demonstrated increased phosphorylation at 120-130 kDa when compared with extracts of intact monolayers. The pp125(FAK) immunoprecipitated from wounded monolayers exhibited increased kinase activity as compared to pp125(FAK) from intact monolayers. The time to wound closure in HUVEC monolayers was doubled by tyrphostin AG 213 treatment. The same concentration of AG 213 interfered with HUVEC focal adhesion and stress fiber formation. AG 213 inhibited adhesion-associated tyrosine phosphorylation of pp125(FAK) in HUVEC. Tyrphostins AG 213 and AG 808 inhibited pp125(FAK) activity in in vitro kinase assays. pp125(FAK) immunoprecipitates from HUVEC treated with both of these inhibitors also had kinase activity in vitro that was below levels seen in untreated HUVEC. These findings suggest that tyrosine phosphorylation of cytoskeletal proteins may be important in HUVEC spreading and migration and that pp125(FAK) may mediate phosphotyrosine formation during these processes.

Original languageEnglish (US)
Pages (from-to)349-361
Number of pages13
JournalMolecular biology of the cell
Volume5
Issue number3
DOIs
StatePublished - Mar 1994
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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