Thrombin stimulates phosphorylation of insulin-like growth factor-1 receptor, insulin receptor substrate-1, and phospholipase C-γ1 in rat aortic smooth muscle cells

Rao Gadiparthi, P. Delafontaine, M. S. Runge

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Abstract

It has recently been reported that protein-tyrosine kinase activity is required for thrombin-induced growth in vascular smooth muscle cells (VSMC). In the present study, we have identified several phosphoproteins that are tyrosine-phosphorylated in response to thrombin in quiescent VSMC. These proteins are insulin-like growth factor-1 receptor β-subunit (IGF-1Rβ), insulin receptor substrate-1 (IRS-1), and phospholipase C-γ1 (PLC-γ1). Thrombin-stimulated phosphorylation of these proteins was rapid; it was maximal at 1 min and reduced thereafter. Thrombin also activated mitogen- activated protein kinases (MAPK) in quiescent VSMC in a biphasic manner with a rapid and larger peak at 10 min (6-fold) followed by a sustained smaller second peak at 2 h (2-fold). Inhibition of protein-tyrosine kinase activity by the use of two structurally different protein-tyrosine kinase inhibitors, genistein and herbimycia A, significantly blocked the thrombin-induced tyrosine phosphorylation of IGF-1Rβ, IRS-1, and PLC-γ1 and decreased thrombin-stimulated DNA synthesis. In contrast, however, inhibition of protein-tyrosine kinase activity had no effect on thrombin activation of MAPK. Collectively, these findings suggest a role for tyrosine phosphorylation of IGF-1Rβ, IRS-1, and PLC-γ1 in thrombin-induced mitogenic signaling events in VSMC. Furthermore, while protein tyrosine phosphorylation is essential for thrombin-induced DNA synthesis, it is not required for thrombin-stimulated MAPK activation. Since thrombin rapidly activated Src in VSMC, Src may be involved in the cross-talk between the G-protein-coupled receptor agonist and a tyrosine kinase receptor such as IGF-1R.

Original languageEnglish (US)
Pages (from-to)27871-27875
Number of pages5
JournalJournal of Biological Chemistry
Volume270
Issue number46
DOIs
StatePublished - Jan 1 1995

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Somatomedin Receptors
Insulin Receptor Substrate Proteins
Phosphorylation
Type C Phospholipases
Somatomedins
Thrombin
Smooth Muscle Myocytes
Muscle
Rats
Cells
Vascular Smooth Muscle
Protein-Tyrosine Kinases
Tyrosine
Mitogen-Activated Protein Kinases
Chemical activation
Proteins
Genistein
Phosphoproteins
DNA
Receptor Protein-Tyrosine Kinases

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

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title = "Thrombin stimulates phosphorylation of insulin-like growth factor-1 receptor, insulin receptor substrate-1, and phospholipase C-γ1 in rat aortic smooth muscle cells",
abstract = "It has recently been reported that protein-tyrosine kinase activity is required for thrombin-induced growth in vascular smooth muscle cells (VSMC). In the present study, we have identified several phosphoproteins that are tyrosine-phosphorylated in response to thrombin in quiescent VSMC. These proteins are insulin-like growth factor-1 receptor β-subunit (IGF-1Rβ), insulin receptor substrate-1 (IRS-1), and phospholipase C-γ1 (PLC-γ1). Thrombin-stimulated phosphorylation of these proteins was rapid; it was maximal at 1 min and reduced thereafter. Thrombin also activated mitogen- activated protein kinases (MAPK) in quiescent VSMC in a biphasic manner with a rapid and larger peak at 10 min (6-fold) followed by a sustained smaller second peak at 2 h (2-fold). Inhibition of protein-tyrosine kinase activity by the use of two structurally different protein-tyrosine kinase inhibitors, genistein and herbimycia A, significantly blocked the thrombin-induced tyrosine phosphorylation of IGF-1Rβ, IRS-1, and PLC-γ1 and decreased thrombin-stimulated DNA synthesis. In contrast, however, inhibition of protein-tyrosine kinase activity had no effect on thrombin activation of MAPK. Collectively, these findings suggest a role for tyrosine phosphorylation of IGF-1Rβ, IRS-1, and PLC-γ1 in thrombin-induced mitogenic signaling events in VSMC. Furthermore, while protein tyrosine phosphorylation is essential for thrombin-induced DNA synthesis, it is not required for thrombin-stimulated MAPK activation. Since thrombin rapidly activated Src in VSMC, Src may be involved in the cross-talk between the G-protein-coupled receptor agonist and a tyrosine kinase receptor such as IGF-1R.",
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T1 - Thrombin stimulates phosphorylation of insulin-like growth factor-1 receptor, insulin receptor substrate-1, and phospholipase C-γ1 in rat aortic smooth muscle cells

AU - Gadiparthi, Rao

AU - Delafontaine, P.

AU - Runge, M. S.

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N2 - It has recently been reported that protein-tyrosine kinase activity is required for thrombin-induced growth in vascular smooth muscle cells (VSMC). In the present study, we have identified several phosphoproteins that are tyrosine-phosphorylated in response to thrombin in quiescent VSMC. These proteins are insulin-like growth factor-1 receptor β-subunit (IGF-1Rβ), insulin receptor substrate-1 (IRS-1), and phospholipase C-γ1 (PLC-γ1). Thrombin-stimulated phosphorylation of these proteins was rapid; it was maximal at 1 min and reduced thereafter. Thrombin also activated mitogen- activated protein kinases (MAPK) in quiescent VSMC in a biphasic manner with a rapid and larger peak at 10 min (6-fold) followed by a sustained smaller second peak at 2 h (2-fold). Inhibition of protein-tyrosine kinase activity by the use of two structurally different protein-tyrosine kinase inhibitors, genistein and herbimycia A, significantly blocked the thrombin-induced tyrosine phosphorylation of IGF-1Rβ, IRS-1, and PLC-γ1 and decreased thrombin-stimulated DNA synthesis. In contrast, however, inhibition of protein-tyrosine kinase activity had no effect on thrombin activation of MAPK. Collectively, these findings suggest a role for tyrosine phosphorylation of IGF-1Rβ, IRS-1, and PLC-γ1 in thrombin-induced mitogenic signaling events in VSMC. Furthermore, while protein tyrosine phosphorylation is essential for thrombin-induced DNA synthesis, it is not required for thrombin-stimulated MAPK activation. Since thrombin rapidly activated Src in VSMC, Src may be involved in the cross-talk between the G-protein-coupled receptor agonist and a tyrosine kinase receptor such as IGF-1R.

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