AMINO-TERMINAL SEQUENCE OF P36 AND ASSOCIATED P10 - IDENTIFICATION OF THE SITE OF TYROSINE PHOSPHORYLATION AND HOMOLOGY WITH S-100
AMINO-TERMINAL SEQUENCE OF P36 AND ASSOCIATED P10 - IDENTIFICATION OF THE SITE OF TYROSINE PHOSPHORYLATION AND HOMOLOGY WITH S-100
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DOI：
10.1073/pnas.82.23.7884
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
TACK, BF
中科院分区：
文献类型：
--
作者：
GLENNEY, JR;TACK, BF
p36是病毒和生长因子受体相关酪氨酸蛋白激酶的主要底物。p36可以作为由Mr 36，000的亚基（p36）和Mr 10，000的亚基（p10）组成的复合物分离，并且它代表丰富的细胞蛋白。我们已经从牛肠上皮细胞中分离出p36-p10复合物，并分析了这两个亚基的氨基末端。p10的前56个氨基酸的序列分析表明与牛脑中的Mr 10，000钙结合蛋白（称为S-100）具有惊人的序列同源性（48%相同的位置残基）。肠p36可以在体外用免疫沉淀的pp 60 v-src和[γ-Src]有效地标记在单个酪氨酸上。[32 P]ATP。p36与糜蛋白酶的温和蛋白水解导致裂解成大（Mr，33，000）和小域（Mr，3000），后者代表磷酸化的氨基末端。虽然氨基末端明显被阻断，但Mr 3000片段的二级胰蛋白酶肽以及Mr 33，000结构域和重叠肽的氨基末端序列的序列分析清楚地确定了酪氨酸磷酸化的位点。
p36 is a major substrate of both viral and growth factor-receptor-associated tyrosine protein kinases. p36 can be isolated as a complex consisting of a subunit of Mr 36,000 (p36) and a subunit of Mr 10,000 (p10), and it represents an abundant cellular protein. We have isolated the p36-p10 complex from bovine intestinal epithelium and analyzed the amino terminus of both subunits. Sequence analysis of the first 56 amino acids of p10 demonstrates a striking sequence homology (48% identically placed residues) with the Mr 10,000 calcium-binding proteins from bovine brain, termed S-100. Intestinal p36 could be effectively labeled on a single tyrosine in vitro with immunoprecipitated pp60v-src and [.gamma.-32P]ATP. Mild proteolysis of p36 with chymotrypsin resulted in the cleavage into large (Mr, 33,000) and small domains (Mr, 3000), with the latter representing the phosphorylated amino terminus. Although the amino terminus is apparently blocked, sequence analysis of a secondary tryptic peptide of the Mr 3000 fragment as well as the amino-terminal sequence of the Mr 33,000 domain and overlapping peptides clearly established the site of tryosine phosphorylation.