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正常细胞和多瘤病毒转化细胞中p59fyn(一种src样蛋白酪氨酸激酶)的鉴定与特性分析

Identification and characterization of p59fyn (a src-like protein tyrosine kinase) in normal and polyoma virus transformed cells.

作者信息

Kypta R M, Hemming A, Courtneidge S A

机构信息

European Molecular Biology Laboratory, Heidelberg, FRG.

出版信息

EMBO J. 1988 Dec 1;7(12):3837-44. doi: 10.1002/j.1460-2075.1988.tb03269.x.

Abstract

fyn is a member of the growing family of protein tyrosine kinase genes whose sequences are highly related to that of c-src. We have generated antibodies to peptides corresponding to two different amino-terminal sequences encoded by this gene. Antisera to both peptides recognized a 59 kd protein from human and mouse fibroblasts. p59fyn was phosphorylated in vivo on serine and tyrosine residues and was also myristylated. Furthermore, immune precipitates of p59fyn had tyrosine kinase activity in vitro, as measured by autophosphorylation and by phosphorylation of substrates such as enolase. This kinase activity was shown to be negatively regulated by tyrosine phosphorylation. We have also established that, like pp60c-src and p62c-yes, p59fyn was complexed with middle T antigen, the transforming protein of polyoma virus. However, the tyrosine kinase activity of p59fyn was not elevated in middle T transformed cells. Possible explanations for this are discussed.

摘要

Fyn是蛋白质酪氨酸激酶基因不断增多的家族中的一员,其序列与c-src的序列高度相关。我们针对该基因编码的两种不同氨基末端序列对应的肽段制备了抗体。针对这两种肽段的抗血清均识别来自人和小鼠成纤维细胞的一种59 kd蛋白。p59fyn在体内丝氨酸和酪氨酸残基上发生磷酸化,并且也进行了肉豆蔻酰化修饰。此外,通过自身磷酸化以及烯醇化酶等底物的磷酸化检测发现,p59fyn的免疫沉淀物在体外具有酪氨酸激酶活性。这种激酶活性被证明受到酪氨酸磷酸化的负调控。我们还证实,与pp60c-src和p62c-yes一样,p59fyn与多瘤病毒的转化蛋白中间T抗原形成复合物。然而,在中间T抗原转化的细胞中,p59fyn的酪氨酸激酶活性并未升高。对此的可能解释进行了讨论。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2578/454962/4075091d6e8d/emboj00149-0208-a.jpg

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