Bogoyevitch M A, Glennon P E, Sugden P H
Department of Cardiac Medicine, National Heart and Lung Institute, University of London, UK.
FEBS Lett. 1993 Feb 15;317(3):271-5. doi: 10.1016/0014-5793(93)81291-7.
ET-1 stimulated MBP kinase activity in cultured cardiomyocytes. Maximal activation (3.5-fold) was at 5 min. EC50 was 0.2 nM. PMA or PE also increased MBP kinase (4- or 2.5-fold, respectively). Pre-treatment with PMA down-regulated the subsequent response to ET-1 or PMA. ET-1- or PMA-stimulated MBP kinase was resolved into 2 major (peaks II and IV) and 2 minor peaks by FPLC on Mono Q. Peaks II and IV were inactivated by either LAR or PP2A. Renatured MBP kinase activities following SDS-PAGE in MBP-containing gels and immunoblot analysis showed that peak II was a p42 MAP kinase and peak IV was a p44 MAP kinase.
内皮素 -1（ET -1）刺激培养的心肌细胞中的髓鞘碱性蛋白（MBP）激酶活性。最大激活（3.5倍）出现在5分钟时。半数有效浓度（EC50）为0.2纳摩尔。佛波酯（PMA）或苯肾上腺素（PE）也增加MBP激酶活性（分别为4倍或2.5倍）。用PMA预处理会下调随后对ET -1或PMA的反应。通过在Mono Q上进行快速蛋白质液相色谱（FPLC）分析，ET -1或PMA刺激的MBP激酶可分离为2个主要峰（峰II和峰IV）和2个次要峰。峰II和峰IV可被淋巴细胞特异性酪氨酸激酶（LAR）或蛋白磷酸酶2A（PP2A）灭活。在含MBP的凝胶中进行十二烷基硫酸钠 -聚丙烯酰胺凝胶电泳（SDS -PAGE）后复性的MBP激酶活性以及免疫印迹分析表明，峰II是p42丝裂原活化蛋白激酶（MAP激酶），峰IV是p44 MAP激酶。
