Whikehart D R, Soppet D R
Invest Ophthalmol Vis Sci. 1981 Dec;21(6):819-25.
Plasma membranes from bovine corneal endothelial cells have been prepared for assay of ion transport enzymes (ATPase, EC 3.6.1.8). The membrane were judged to be highly purified by protein recovery, electron microscopy, and assay of enzyme markers. Sodium-potassium-stimulated ATPase was more than six times more active in the membranes (6.7 mumoles phosphate released/mg protein/30 min) than the whole homogenates (1.1 mumoles phosphate released). However, no increase in activity was seen with anionic ATPase of membranes (2.5 mumoles phosphate released with bicarbonate stimulation) vs. whole homogenates (3.0 mumoles phosphate released; insignificant difference). In contrast, the bicarbonate-stimulated ATPase activity of a mitochondrial preparation from the same cells was 10.6 mumoles phosphate released (p less than 0.001). It is suggested that anionic ATPase has no direct role in deturgescence as part of and endothelial cell "pump."
已制备牛角膜内皮细胞的质膜,用于离子转运酶(ATP酶,EC 3.6.1.8)的测定。通过蛋白质回收率、电子显微镜检查和酶标志物测定,判断这些膜已高度纯化。钠钾刺激的ATP酶在膜中的活性(6.7微摩尔磷酸盐释放/毫克蛋白质/30分钟)比全匀浆(1.1微摩尔磷酸盐释放)高六倍多。然而,膜的阴离子ATP酶(碳酸氢盐刺激下释放2.5微摩尔磷酸盐)与全匀浆(释放3.0微摩尔磷酸盐;差异不显著)相比,活性没有增加。相反,来自相同细胞的线粒体制剂在碳酸氢盐刺激下的ATP酶活性为10.6微摩尔磷酸盐释放(p小于0.001)。有人提出,阴离子ATP酶作为内皮细胞“泵”的一部分,在消肿过程中没有直接作用。