Increased tyrosine kinase activity in pp60c-src immunoprecipitate from platelet activating factor stimulated human platelets: in vitro phosphorylation of a synthetic peptide.
Increased tyrosine kinase activity in pp60c-src immunoprecipitate from platelet activating factor stimulated human platelets: in vitro phosphorylation of a synthetic peptide.
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血小板激活因子刺激的人血小板的 pp60c-src 免疫沉淀物中酪氨酸激酶活性增加：合成肽的体外磷酸化。
DOI：
10.1016/0024-3205(93)90665-p
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发表时间：
1993
期刊：
影响因子：
6.1
通讯作者：
Shukla,SD
中科院分区：
文献类型：
--
作者：
Zhu,CY;Shukla,SD
用血小板活化因子（PAF）刺激人血小板，研究pp 60 c-src酪氨酸激酶的参与。血小板pp 60 c-src免疫沉淀后，用pp 60 v-src单克隆抗体免疫印迹，经酶谱网检测，显示60、50和29 kDa四条蛋白带。在PAF刺激的血小板pp 60 c-src免疫沉淀物中，这些条带的磷酸化程度增加。为了测定酪氨酸激酶活性，我们使用了13个氨基酸的合成肽（Arg → Arg → Leu → Ile → Glu → Asp → Ala → Glu → Tyr → Ala → Arg →;Gly），其含有与pp 60 c-src上的磷酸化位点相似的序列。pp 60 c-src免疫沉淀物与肽和[32 P]ATP孵育引起该肽在体外磷酸化。当使用正常小鼠IgG结合蛋白代替pp 60 c-src免疫沉淀物时，未观察到这种肽磷酸化。血小板活化因子处理后的pp 60 c-src免疫沉淀物可明显增加血小板肽的磷酸化。Lyso-PAF对磷酸化无影响。PAF拮抗剂CV-6209和WEB-2086阻断PAF刺激的磷酸化。这表明这种反应具有结构特异性和PAF受体依赖性。这些结果为PAF刺激人血小板增加pp 60 c-src免疫沉淀物中酪氨酸激酶活性提供了直接证据。
The involvement of pp60c-srctyrosine kinase was studied in human platelets stimulated with platelet activating factor (PAF). Immunoprecipitation of pp60c-srcfrom platelets followed by immunoblot with pp60v-srcmonoclonal antibody revealed four protein bands of 60, 50 and 29 kDa as detected by enzymographic web. The phosphorylation of these bands was increased in the pp60c-srcimmunoprecipitate from PAF stimulated platelets. To assay the tyrosine kinase activity, we used a 13 amino acid synthetic peptide (ArgArgLeuIleGluAspAlaGluTyrAlaArg;Gly) which contains sequences similar to the phosphorylation site on pp60c-src. Incubation of the pp60c-srcimmunoprecipitate with the peptide and [32P]ATP caused phosphorylation of this peptidein vitro. This peptide phosphorylation was not observed when normal mouse IgG-bound protein(s) was used instead of pp60c-srcimmunoprecipitate. The peptide phosphorylation was markedly increased by pp60c-srcimmunoprecipitate obtained from PAF treated platelets. Lyso-PAF had no effect on the phosphorylation. PAF antagonists CV-6209 and WEB-2086 blocked PAF stimulated phosphorylation. This indicated structurally specific and PAF receptor dependency of this response. These results provide direct evidence that PAF stimulation of human platelets increased tyrosine kinase activity in pp60c-srcimmunoprecipitate.
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酪氨酸激酶的血小板激活因子刺激及其与兔血小板中磷脂酶 C 的关系：金雀异黄素和磷酸酪氨酸单克隆抗体的研究。
影响因子：
3.6
作者：
Dhar,A;Paul,AK;Shukla,SD
通讯作者：
Shukla,SD
厄布他汀阻断血小板激活因子诱导的蛋白酪氨酸磷酸化、多磷酸肌醇水解、蛋白激酶 C 激活、血清素分泌和兔血小板聚集
影响因子：
3.5
作者：
H. Salari;V. Duronio;Sandra L. Howard;M. Demos;Kelvin Jones;A. Reany;A. T. Hudson;S. Pelech
通讯作者：
S. Pelech
鉴定与 pp60c-src 氨基末端肽相互作用的血小板膜蛋白。
DOI：
--
发表时间：
1991
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Feder,D;Bishop,JM
通讯作者：
Bishop,JM
影响因子：
5.4
作者：
SEFTON, BM;HUNTER, T;BEEMON, K
通讯作者：
BEEMON, K
正常非增殖细胞中的高酪氨酸激酶活性
影响因子：
64.8
作者：
F. Tuy;J. Henry;C. Rosenfeld;A. Kahn
通讯作者：
A. Kahn