Bradykinin-stimulated protein tyrosine phosphorylation promotes endothelial nitric oxide synthase translocation to the cytoskeleton.
Bradykinin-stimulated protein tyrosine phosphorylation promotes endothelial nitric oxide synthase translocation to the cytoskeleton.
复制标题
DOI：
10.1006/bbrc.1996.1417
复制
发表时间：
1996-09
影响因子：
3.1
通讯作者：
V. Venema;Mario B. Marrero;R. Venema
中科院分区：
文献类型：
--
作者：
V. Venema;Mario B. Marrero;R. Venema
用缓激肽刺激牛主动脉内皮细胞（BAEC）产生90 kDa内皮型一氧化氮合酶（eNOS）相关蛋白的酪氨酸磷酸化/去磷酸化循环，我们将其称为ENAP-1（内皮型一氧化氮合酶相关蛋白1）。ENAP-1与BAEC中的eNOS特异性且紧密地相互作用，并且与抗eNOS抗体从细胞裂解物中共免疫沉淀。此外，抗磷酸酪氨酸抗体会共沉淀eNOS。缓激肽刺激的ENAP-1的酪氨酸磷酸化被酪氨酸激酶抑制剂tyrphostin阻断。去磷酸化被酪氨酸磷酸酶抑制剂原钒酸盐阻断。用缓激肽或酪氨酸磷酸酶抑制剂氧化苯胂治疗BAEC可促进洗涤剂不溶性细胞骨架蛋白的酪氨酸磷酸化，并伴有eNOS易位至细胞骨架亚细胞区室。易位被酪氨酸激酶抑制剂格尔德霉素阻断，并且似乎不改变酶催化活性。eNOS与细胞骨架的酪氨酸磷酸化依赖性关联可能在将NO产生靶向至特定亚细胞位置中具有作用。
Stimulation of bovine aortic endothelial cells (BAEC) with bradykinin produces cycles of tyrosine phosphorylation/dephosphorylation of a 90 kDa endothelial nitric oxide synthase (eNOS)-associated protein which we have termed ENAP-1 (for endothelial nitric oxide synthase-associated protein 1). ENAP-1 interacts specifically and tightly with eNOS in BAEC and is co-immunoprecipitated from cell lysates with anti-eNOS antibodies. In addition, anti-phosphotyrosine antibodies co-precipitate eNOS. Bradykinin-stimulated tyrosine phosphorylation of ENAP-1 is blocked by the tyrosine kinase inhibitor, tyrphostin. Dephosphorylation is blocked by the tyrosine phosphatase inhibitor, orthovanadate. Treatment of BAEC with bradykinin or the tyrosine phosphatase inhibitor, phenylarsine oxide promotes tyrosine phosphorylation of detergent-insoluble, cytoskeletal proteins accompanied by translocation of eNOS to the cytoskeletal subcellular compartment. Translocation is blocked by the tyrosine kinase inhibitor, geldanamycin and does not appear to alter enzyme catalytic activity. Tyrosine phosphorylation-dependent association of eNOS with the cytoskeleton may have a role in targeting NO production to specific subcellular locations.