Membranes from T and B lymphocytes have different patterns of tyrosine phosphorylation.
Membranes from T and B lymphocytes have different patterns of tyrosine phosphorylation.
复制标题
T 和 B 淋巴细胞的膜具有不同的酪氨酸磷酸化模式。
DOI：
10.1073/pnas.81.8.2347
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发表时间：
1984
影响因子：
11.1
通讯作者：
Gillespie,GY
中科院分区：
文献类型：
--
作者：
Earp,HS;Austin,KS;Buessow,SC;Dy,R;Gillespie,GY
从小鼠和大鼠脾脏分离的膜组分表达大量酪氨酸特异性蛋白激酶活性。磷酸酪氨酸（P-Tyr）的积累在内源性膜基质的钒酸盐或非离子去污剂刺激。当在0 ℃下在1 mM Mn 2+和Triton X-100存在下进行体外磷酸化时，P-Tyr占总磷酸化氨基酸的40-50%。聚丙烯酰胺凝胶电泳显示，混合淋巴细胞群体的膜有四个主要的含P-Tyr的蛋白质。而非离子洗涤剂是有效的刺激P-Tyr积累在所有四个基板，其中两个（p61和p55）的酪氨酸磷酸化显着依赖于钒酸盐。这两种底物存在于通过亲和色谱法纯化的表面Ig-承载脾淋巴细胞和Raji（一种人B-淋巴母细胞样细胞系）的膜中。在脾细胞膜（p64和p58）中观察到的其他两种底物中的P-Tyr蓄积对钒酸盐的依赖性要小得多。p64和p58在小鼠胸腺细胞和人及小鼠T淋巴瘤细胞系的膜中磷酸化，而p61和p55没有。因此，似乎在鼠和人淋巴细胞中，p64和p58充当T细胞特异性底物，而p61和p55特异性地与B淋巴细胞相关。此外，这些不同的P-Tyr底物模式在源自B和T细胞的一些肿瘤细胞系中是保守的。
Membrane fractions isolated from mouse and rat spleen expressed substantial tyrosine-specific protein kinase activity. Phosphotyrosine (P-Tyr) accumulation in endogenous membrane substrates was stimulated by vanadate or nonionic detergents. When in vitro phosphorylation was carried out at 0 degree C in the presence of 1 mM Mn2+ and Triton X-100, P-Tyr constituted up to 40-50% of the total phospho amino acid. Polyacrylamide gel electrophoresis showed that membranes from mixed lymphocyte populations have four major P-Tyr-containing proteins. Whereas nonionic detergents were potent stimuli for P-Tyr accumulation in all four substrates, tyrosine phosphorylation of two of these (p61 and p55) was markedly dependent on vanadate. These two substrates were present in membranes from surface Ig-bearing splenic lymphocytes purified by affinity chromatography and Raji, a human B-lymphoblastoid cell line. P-Tyr accumulation in the two other substrates observed in splenocyte membranes (p64 and p58) was much less dependent on vanadate. p64 and p58 were phosphorylated in membranes from mouse thymocytes and human and mouse T-lymphoma cell lines, while p61 and p55 were not. Thus it appears that in both murine and human lymphocytes, p64 and p58 served as T-cell-specific substrates, while p61 and p55 were specifically associated with B lymphocytes. Moreover, these distinct P-Tyr substrate patterns were conserved in some neoplastic cell lines derived from B and T cells.