Bombesin, Bradykinin, Vasopressin, and Phorbol Esters Rapidly and Transiently Activate Src Family Tyrosine Kinases in Swiss 3T3 Cells

Bombesin, Bradykinin, Vasopressin, and Phorbol Esters Rapidly and Transiently Activate Src Family Tyrosine Kinases in Swiss 3T3 Cells
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铃蟾肽、缓激肽、加压素和佛波酯快速瞬时激活 Swiss 3T3 细胞中的 Src 家族酪氨酸激酶

DOI:
--
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发表时间:
1996
影响因子:
4.8
通讯作者:
E. Rozengurt
E. Rozengurt
中科院分区:
生物学2区
文献类型:
--
作者:
JoséLuis Rodriguez;E. Rozengurt

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通过免疫复合物激酶试验中的自磷酸化(4.6 ± 0.2倍刺激,n = 44)和外源性底物的磷酸化测定,用蛙皮素处理静止的Swiss 3 T3细胞可诱导Src家族酪氨酸激酶活性的快速(≤40 s)和短暂增加。佛波醇12,13-二丁酸酯以类似的动力学增加Src家族激酶的活性,但不如蛙皮素有效。然而,蛙皮素激活Src家族激酶不依赖于蛋白激酶C或Ca 2+。蛙皮素刺激Src家族激酶活性也可以与p125粘着斑激酶酪氨酸磷酸化分离。无论是治疗细胞松弛素D,也不放置在悬浮液中的细胞防止刺激的Src家族激酶活性诱导的蛙皮素,但都废除蛙皮素诱导的p125粘着斑激酶的酪氨酸磷酸化。蛙皮素对Src家族激酶活性的刺激被钒酸盐(一种有效的蛋白酪氨酸磷酸酶抑制剂)完全阻止。缓激肽和加压素也刺激Src家族激酶活性短暂,这种刺激也被钒酸盐抑制。我们的研究结果剖析了两个独立的途径,导致蛋白质酪氨酸磷酸化的神经肽刺激瑞士3 T3细胞。
Treatment of quiescent Swiss 3T3 cells with bombesin induces a rapid (≤40 s) and transient increase in the kinase activity of the Src family of tyrosine kinases, as determined by autophosphorylation in immune complex kinase assays (4.6 ± 0.2-fold stimulation, n = 44) and phosphorylation of exogenous substrates. Phorbol 12,13-dibutyrate increased the activity of Src family kinases with similar kinetics but was less effective than bombesin. However, Src family kinase activation by bombesin is not dependent either on protein kinase C or Ca2+. Bombesin stimulation of Src family kinase activity could also be dissociated from p125 focal adhesion kinase tyrosine phosphorylation. Neither treatment with cytochalasin D nor placement of the cells in suspension prevented the stimulation of Src family kinase activity induced by bombesin, but both abolished bombesin-induced tyrosine phosphorylation of p125 focal adhesion kinase. The stimulation of the Src family kinase activity by bombesin was completely prevented by treatment with vanadate, a potent inhibitor of protein-tyrosine phosphatases. Bradykinin and vasopressin also stimulated Src family kinase activity transiently, and this stimulation was also inhibited by vanadate. Our results dissect two separate pathways that lead to protein tyrosine phosphorylation in neuropeptide-stimulated Swiss 3T3 cells.
以 pp60c-src 羧基末端为模型的固定化合成磷酸肽选择性结合激活的 pp60c-src。
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DOI: --
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