The tyrosine kinase p56lck mediates activation of swelling-induced chloride channels in lymphocytes.
The tyrosine kinase p56lck mediates activation of swelling-induced chloride channels in lymphocytes.
复制标题
酪氨酸激酶p56LCK介导淋巴细胞中溶胀诱导的氯化物通道的激活。
DOI：
10.1083/jcb.141.1.281
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发表时间：
1998-04-06
期刊：
影响因子：
--
通讯作者：
Lang F
中科院分区：
文献类型：
--
作者：
Lepple-Wienhues A;Szabò I;Laun T;Kaba NK;Gulbins E;Lang F
渗透性细胞膨胀激活Cl -通道，实现阴离子外排。在这项研究中，我们发现酪氨酸激酶抑制剂herbimycin A和基因敲除p56lck（一种src样酪氨酸激酶）可以阻断人T细胞系的调节体积减少（regulatory volume减少，RVD）。在全细胞膜片钳实验中，溶胀激活的氯离子电流（ICl - swell）被herbimycin a和lavendustin阻断。在p56lck缺陷细胞中，ICl−swell的渗透激活是有缺陷的。p56lck的再转染恢复渗透电流激活。此外，酪氨酸激酶活性足以激活ICl - swell。将纯化的p56lck添加到切除的斑块中，激活具有31 pS单位电导的向外整流氯离子通道。纯化的p56lck洗涤到细胞质中，即使在低渗的细胞内溶液导致细胞收缩时，也能激活原生细胞和p56lck缺陷细胞的ICl -膨胀。当全细胞电流被肿胀或p56lck激活时，可以观察到缓慢的单通道门控事件，显示25-28 pS的统一电导。根据我们的膜片钳数据，渗透肿胀增加了免疫沉淀的p56lck的活性。我们得出结论，渗透性肿胀通过酪氨酸激酶p56lck激活淋巴细胞的ICl -肿胀。
Osmotic cell swelling activates Cl− channels to achieve anion efflux. In this study, we find that both the tyrosine kinase inhibitor herbimycin A and genetic knockout of p56lck, a src-like tyrosine kinase, block regulatory volume decrease (RVD) in a human T cell line. Activation of a swelling-activated chloride current (ICl−swell) by osmotic swelling in whole-cell patch-clamp experiments is blocked by herbimycin A and lavendustin. Osmotic activation of ICl−swell is defective in p56lck-deficient cells. Retransfection of p56lck restores osmotic current activation. Furthermore, tyrosine kinase activity is sufficient for activation of ICl−swell. Addition of purified p56lck to excised patches activates an outwardly rectifying chloride channel with 31 pS unitary conductance. Purified p56lck washed into the cytoplasm activates ICl−swell in native and p56lck-deficient cells even when hypotonic intracellular solutions lead to cell shrinkage. When whole-cell currents are activated either by swelling or by p56lck, slow single-channel gating events can be observed revealing a unitary conductance of 25–28 pS. In accordance with our patch-clamp data, osmotic swelling increases activity of immunoprecipitated p56lck. We conclude that osmotic swelling activates ICl−swell in lymphocytes via the tyrosine kinase p56lck.