Tyrosine phosphorylation and bradykinin-induced signaling in endothelial cells.
It is generally accepted that in endothelial cells the occupation of bradykinin B2 receptors, which are linked to the guanine nucleotide-dependent regulatory proteins, Gi and Gq, results in the activation of phospholipase C-beta1 (PLC-beta1), followed by a transient increase in the formation of inositol 1,4,5-trisphosphate (IP3) and diacylglycerol. The PLC-beta1 isoform, in contrast to the gamma1 isoform, is present only at a low level in cultured endothelial cells, implying that PLC-gamma1 activation may play an important role in endothelial signaling pathways. In cultured human endothelial cells, bradykinin induced a rapid increase in the tyrosine phosphorylation of several Triton-soluble proteins. Immunoprecipitation of tyrosine-phosphorylated proteins from bradykinin-stimulated cells followed by Western blotting using the respective antibodies facilitated the identification of a 77 kiloDalton (kDa) protein as paxillin, a 130 kDa protein as PLC-gamma1, and a 42/44 kDa doublet as mitogen-activated protein (MAP) kinase. The bradykinin-induced tyrosine phosphorylation of PLC-gamma1 was relatively transient and was associated with an increase in intracellular levels of IP3. Bradykinin also induced the rapid and transient activation of phosphotyrosine phosphatases localized mainly in the Triton X-100-soluble cell fraction; this tyrosine phosphatase activity was apparently initiated after the release of Ca2+ from intracellular stores.
Fleming
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I
Zentrum der Physiologie, Klinikum der J.W. Goethe-Universitat, Frankfurt/Main, Germany.
Busse
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R
D000704:Analysis of Variance; D015153:Blotting, Western; D001920:Bradykinin / Q000502:physiology*; D015220:Calcium Channels / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D010766:Phosphorylation; D011233:Precipitin Tests; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D015398:Signal Transduction; D014443:Tyrosine / Q000378:metabolism*; D014471:Umbilical Veins
D015220:Calcium Channels; D014443:Tyrosine; D015544:Inositol 1,4,5-Trisphosphate; D019001:Nitric Oxide Synthase; D011505:Protein-Tyrosine Kinases; D017027:Protein Tyrosine Phosphatases; D001920:Bradykinin