Tyrosine Kinases Regulate the Cytoskeletal Attachment of Integrin αIIbβ3 (Platelet Glycoprotein IIb/IIIa) and the Cellular Retraction of Fibrin Polymers

Simone M. Schoenwaelder, Shaun P. Jackson, Yuping Yuan, Melvena S. Teasdale, Hatem H. Salem, Christina A. Mitchell

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    Abstract

    Integrins promote cell-substratum and cell-cell adhesion by acting as transmembrane linker molecules between extracellular adhesion proteins and the actin-rich cytoskeleton. The integrin αIIbβ3 (platelet glycoprotein IIb/IIIa) is essential for platelet spreading, aggregation, fibrin clot retraction, and for the transduction of extracellular signals. We examined the effect of the specific tyrosine kinase inhibitor herbimycin A on integrin and cytoskeletal-mediated events in thrombin-stimulated platelets. Incubation of washed platelets for 24 h with herbimycin A (5 μM) abolished the thrombin-stimulated cytoskeletal enzyme activity of pp60c-src in parallel with a reduction in the tyrosine phosphorylation of multiple platelet proteins, as assessed with anti-phosphotyrosine immunoblots. However, thrombin-induced activation of protein kinase C and the production of thromboxane A2 were not altered by herbimycin A. Despite the absence of cytoskeletal pp60c-src enzyme activity, platelet shape change, aggregation, and serotonin release were unaltered following platelet stimulation with thrombin (0.05-1.0 unit/ml). Herbimycin A-treated platelets also demonstrated normal platelet aggregation in response to collagen (5 μg/ml), ionophore A23187 (2 μM), and ADP/ adrenaline (10 μM each). However, the ability of herbimycin A-treated platelets to retract fibrin gels was significantly reduced. This defect in clot retraction was associated with reduced incorporation of integrin αIIbβ3 into the cytoskeletal fraction of thrombin-aggregated platelets. Our studies suggest that tyrosine kinases in platelets regulate the cytoskeletal attachment of αIIbβ3, as an essential process for the transmission of cellular contractile forces to fibrin polymers.

    Original languageEnglish
    Pages (from-to)32479-32487
    Number of pages9
    JournalJournal of Biological Chemistry
    Volume269
    Issue number51
    Publication statusPublished - 23 Dec 1994

    Cite this

    @article{304d4f68e5e54d13a7bc5b83da6f9141,
    title = "Tyrosine Kinases Regulate the Cytoskeletal Attachment of Integrin αIIbβ3 (Platelet Glycoprotein IIb/IIIa) and the Cellular Retraction of Fibrin Polymers",
    abstract = "Integrins promote cell-substratum and cell-cell adhesion by acting as transmembrane linker molecules between extracellular adhesion proteins and the actin-rich cytoskeleton. The integrin αIIbβ3 (platelet glycoprotein IIb/IIIa) is essential for platelet spreading, aggregation, fibrin clot retraction, and for the transduction of extracellular signals. We examined the effect of the specific tyrosine kinase inhibitor herbimycin A on integrin and cytoskeletal-mediated events in thrombin-stimulated platelets. Incubation of washed platelets for 24 h with herbimycin A (5 μM) abolished the thrombin-stimulated cytoskeletal enzyme activity of pp60c-src in parallel with a reduction in the tyrosine phosphorylation of multiple platelet proteins, as assessed with anti-phosphotyrosine immunoblots. However, thrombin-induced activation of protein kinase C and the production of thromboxane A2 were not altered by herbimycin A. Despite the absence of cytoskeletal pp60c-src enzyme activity, platelet shape change, aggregation, and serotonin release were unaltered following platelet stimulation with thrombin (0.05-1.0 unit/ml). Herbimycin A-treated platelets also demonstrated normal platelet aggregation in response to collagen (5 μg/ml), ionophore A23187 (2 μM), and ADP/ adrenaline (10 μM each). However, the ability of herbimycin A-treated platelets to retract fibrin gels was significantly reduced. This defect in clot retraction was associated with reduced incorporation of integrin αIIbβ3 into the cytoskeletal fraction of thrombin-aggregated platelets. Our studies suggest that tyrosine kinases in platelets regulate the cytoskeletal attachment of αIIbβ3, as an essential process for the transmission of cellular contractile forces to fibrin polymers.",
    author = "Schoenwaelder, {Simone M.} and Jackson, {Shaun P.} and Yuping Yuan and Teasdale, {Melvena S.} and Salem, {Hatem H.} and Mitchell, {Christina A.}",
    year = "1994",
    month = "12",
    day = "23",
    language = "English",
    volume = "269",
    pages = "32479--32487",
    journal = "Journal of Biological Chemistry",
    issn = "1083-351X",
    publisher = "American Society for Biochemistry and Molecular Biology",
    number = "51",

    }

    Tyrosine Kinases Regulate the Cytoskeletal Attachment of Integrin αIIbβ3 (Platelet Glycoprotein IIb/IIIa) and the Cellular Retraction of Fibrin Polymers. / Schoenwaelder, Simone M.; Jackson, Shaun P.; Yuan, Yuping; Teasdale, Melvena S.; Salem, Hatem H.; Mitchell, Christina A.

    In: Journal of Biological Chemistry, Vol. 269, No. 51, 23.12.1994, p. 32479-32487.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - Tyrosine Kinases Regulate the Cytoskeletal Attachment of Integrin αIIbβ3 (Platelet Glycoprotein IIb/IIIa) and the Cellular Retraction of Fibrin Polymers

    AU - Schoenwaelder, Simone M.

    AU - Jackson, Shaun P.

    AU - Yuan, Yuping

    AU - Teasdale, Melvena S.

    AU - Salem, Hatem H.

    AU - Mitchell, Christina A.

    PY - 1994/12/23

    Y1 - 1994/12/23

    N2 - Integrins promote cell-substratum and cell-cell adhesion by acting as transmembrane linker molecules between extracellular adhesion proteins and the actin-rich cytoskeleton. The integrin αIIbβ3 (platelet glycoprotein IIb/IIIa) is essential for platelet spreading, aggregation, fibrin clot retraction, and for the transduction of extracellular signals. We examined the effect of the specific tyrosine kinase inhibitor herbimycin A on integrin and cytoskeletal-mediated events in thrombin-stimulated platelets. Incubation of washed platelets for 24 h with herbimycin A (5 μM) abolished the thrombin-stimulated cytoskeletal enzyme activity of pp60c-src in parallel with a reduction in the tyrosine phosphorylation of multiple platelet proteins, as assessed with anti-phosphotyrosine immunoblots. However, thrombin-induced activation of protein kinase C and the production of thromboxane A2 were not altered by herbimycin A. Despite the absence of cytoskeletal pp60c-src enzyme activity, platelet shape change, aggregation, and serotonin release were unaltered following platelet stimulation with thrombin (0.05-1.0 unit/ml). Herbimycin A-treated platelets also demonstrated normal platelet aggregation in response to collagen (5 μg/ml), ionophore A23187 (2 μM), and ADP/ adrenaline (10 μM each). However, the ability of herbimycin A-treated platelets to retract fibrin gels was significantly reduced. This defect in clot retraction was associated with reduced incorporation of integrin αIIbβ3 into the cytoskeletal fraction of thrombin-aggregated platelets. Our studies suggest that tyrosine kinases in platelets regulate the cytoskeletal attachment of αIIbβ3, as an essential process for the transmission of cellular contractile forces to fibrin polymers.

    AB - Integrins promote cell-substratum and cell-cell adhesion by acting as transmembrane linker molecules between extracellular adhesion proteins and the actin-rich cytoskeleton. The integrin αIIbβ3 (platelet glycoprotein IIb/IIIa) is essential for platelet spreading, aggregation, fibrin clot retraction, and for the transduction of extracellular signals. We examined the effect of the specific tyrosine kinase inhibitor herbimycin A on integrin and cytoskeletal-mediated events in thrombin-stimulated platelets. Incubation of washed platelets for 24 h with herbimycin A (5 μM) abolished the thrombin-stimulated cytoskeletal enzyme activity of pp60c-src in parallel with a reduction in the tyrosine phosphorylation of multiple platelet proteins, as assessed with anti-phosphotyrosine immunoblots. However, thrombin-induced activation of protein kinase C and the production of thromboxane A2 were not altered by herbimycin A. Despite the absence of cytoskeletal pp60c-src enzyme activity, platelet shape change, aggregation, and serotonin release were unaltered following platelet stimulation with thrombin (0.05-1.0 unit/ml). Herbimycin A-treated platelets also demonstrated normal platelet aggregation in response to collagen (5 μg/ml), ionophore A23187 (2 μM), and ADP/ adrenaline (10 μM each). However, the ability of herbimycin A-treated platelets to retract fibrin gels was significantly reduced. This defect in clot retraction was associated with reduced incorporation of integrin αIIbβ3 into the cytoskeletal fraction of thrombin-aggregated platelets. Our studies suggest that tyrosine kinases in platelets regulate the cytoskeletal attachment of αIIbβ3, as an essential process for the transmission of cellular contractile forces to fibrin polymers.

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    M3 - Article

    VL - 269

    SP - 32479

    EP - 32487

    JO - Journal of Biological Chemistry

    JF - Journal of Biological Chemistry

    SN - 1083-351X

    IS - 51

    ER -