Expression of the catalytic domain of myosin light chain kinase increases paracellular permeability
Expression of the catalytic domain of myosin light chain kinase increases paracellular permeability
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DOI：
10.1152/ajpcell.1996.271.5.c1678
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发表时间：
1996-11-01
影响因子：
5.5
通讯作者：
deLanerolle, P
中科院分区：
文献类型：
--
作者：
Hecht, G;Pestic, L;deLanerolle, P
20-kDa 肌球蛋白轻链 (MLC20) 磷酸化引起的收缩事件与上皮紧密连接通透性的调节有关。为了解决这个问题，Madin-Darby 犬肾细胞被鼠白血病逆转录病毒载体转染，该载体含有编码肌球蛋白轻链激酶 (tMK) 催化结构域或 β-半乳糖苷酶基因 (β-gal) 的 DNA。对 P-32(i) 标记的转染细胞免疫沉淀的肌球蛋白进行放射自显影照片分析表明，与表达 β-gal 的细胞相比，表达 tMK 的细胞中 MLC20 磷酸化增加了 3.1 +/- 0.9 倍。磷酸肽图谱证实肌球蛋白轻链激酶导致 MLC20 磷酸化增加。 tMK 细胞单层的跨上皮电阻（屏障功能的测量）一致
Contractile events resulting from phosphorylation of the 20-kDa myosin light chain (MLC20) have been implicated in the regulation of epithelial tight junction permeability. To address this question, Madin-Darby canine kidney cells were transfected with a murine leukemia retroviral vector containing DNA encoding either the catalytic domain of myosin light chain kinase (tMK) or the beta-galactosidase gene (beta-gal). Autoradiograms of sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of myosin immunoprecipitated from P-32(i)-labeled transfected cells demonstrated that MLC20 phosphorylation was increased 3.1 +/- 0.9-fold in cells expressing tMK compared with cells expressing beta-gal. Phosphopeptide mapping confirmed that myosin light chain kinase was responsible for the increased MLC20 phosphorylation. Transepithelial electrical resistance, a measurement of barrier function, of tMK cell monolayers was consistently