Nerve growth factor rapidly stimulates tyrosine phosphorylation of phospholipase C-gamma 1 by a kinase activity associated with the product of the trk protooncogene.
Nerve growth factor rapidly stimulates tyrosine phosphorylation of phospholipase C-gamma 1 by a kinase activity associated with the product of the trk protooncogene.
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神经生长因子通过与 trk 原癌基因产物相关的激酶活性快速刺激磷脂酶 C-gamma 1 的酪氨酸磷酸化。
DOI：
10.1073/pnas.88.13.5650
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发表时间：
1991
影响因子：
11.1
通讯作者：
Kaplan,DR
中科院分区：
文献类型：
--
作者：
Vetter,ML;Martin-Zanca,D;Parada,LF;Bishop,JM;Kaplan,DR
神经生长因子（NGF）促进特定神经元群体的存活和分化。细胞对NGF反应的分子机制知之甚少，但最近出现了两条线索。首先，NGF快速刺激NGF响应性嗜铬细胞瘤细胞系PC 12中几种未鉴定蛋白的酪氨酸磷酸化[Maher，P.（1988）Proc.Natl.Acad.Sci. Acad. Sci. USA 85，6788-6791]。第二，由原癌基因trk（p140 trk）编码的蛋白酪氨酸激酶（酪氨酸激酶受体类别的成员）在NGF处理PC 12细胞后被激活并在酪氨酸上磷酸化[Kaplan，D. R.，Martin-Zanca，D. & Parada，L. F.（1991）Nature（伦敦）350，158-160]。我们现在报告说，神经生长因子迅速诱导磷脂酶C-γ 1（PLC-γ 1）的酪氨酸磷酸化，我们目前的证据表明，负责酪氨酸激酶是p140 trk或密切相关的蛋白质。用NGF处理应答细胞引起PLC-γ 1在酪氨酸和丝氨酸上的磷酸化。PLC-γ 1免疫沉淀从神经生长因子刺激的细胞在体外磷酸化共沉淀蛋白激酶活性，和磷酸化主要发生在酪氨酸。负责的激酶可以从细胞裂解物中耗尽的p140 trk特异性抗体。该程序还耗尽了通常与PLC-γ 1共沉淀的140-kDa蛋白质，并在体内响应于NGF而在酪氨酸上磷酸化。从PLC-γ 1胰蛋白酶肽的分析表明，在体外磷酸化的p140 trk相关的激酶活性的残基在很大程度上与那些在体内磷酸化后，神经生长因子治疗一致。我们的研究结果确定PLC-γ 1作为一个可能的底物的trk编码的酪氨酸激酶，他们提供了一个链接之间的神经生长因子依赖性激活p140 trk和刺激细胞内第二信使途径。
Nerve growth factor (NGF) promotes the survival and differentiation of specific populations of neurons. The molecular mechanisms by which cells respond to NGF are poorly understood, but two clues have emerged recently. First, NGF rapidly stimulates tyrosine phosphorylation of several unidentified proteins in the NGF-responsive pheochromocytoma cell line PC12 [Maher, P. (1988) Proc. Natl. Acad. Sci. USA 85, 6788-6791]. Second, the protein-tyrosine kinase encoded by the protooncogene trk (p140trk), a member of the receptor class of tyrosine kinases, becomes activated and phosphorylated on tyrosine after NGF treatment of PC12 cells [Kaplan, D. R., Martin-Zanca, D. & Parada, L. F. (1991) Nature (London) 350, 158-160]. We now report that NGF rapidly induces tyrosine phosphorylation of phospholipase C-gamma 1 (PLC-gamma 1), and we present evidence that the responsible tyrosine kinase is either p140trk or a closely associated protein. Treatment of responsive cells with NGF elicited phosphorylation of PLC-gamma 1 on tyrosine and serine. PLC-gamma 1 immunoprecipitated from NGF-stimulated cells was phosphorylated in vitro by coprecipitating protein kinase activity, and the phosphorylations occurred principally on tyrosine. The responsible kinase could be depleted from cellular lysates by antibodies specific for p140trk. This procedure also depleted a 140-kDa protein that normally coprecipitated with PLC-gamma 1 and became phosphorylated on tyrosine in vivo in response to NGF. Analysis of tryptic peptides from PLC-gamma 1 indicated that the residues phosphorylated in vitro by p140trk-associated kinase activity were largely congruent with those phosphorylated in vivo after NGF treatment. Our findings identify PLC-gamma 1 as a likely substrate for the trk-encoded tyrosine kinase, and they provide a link between NGF-dependent activation of p140trk and the stimulation of intracellular second messenger pathways.