Preparation of renal cortex basal-lateral and bursh border membranes. Localization of adenylate cyclase and guanylate cyclase activities.
Preparation of renal cortex basal-lateral and bursh border membranes. Localization of adenylate cyclase and guanylate cyclase activities.
复制标题
肾皮质基底外侧膜和刷状缘膜的制备。
DOI：
10.1016/0005-2736(77)90340-6
复制
发表时间：
1977
期刊：
影响因子：
--
通讯作者：
B. Sacktor
中科院分区：
文献类型：
--
作者：
C. Liang;B. Sacktor
制备来自同一肾皮质的质膜的管腔刷状缘和对管腔基底外侧段。刷状缘膜富含海藻糖酶和γ-谷氨酰转肽酶，而基底-侧膜富含（Na++ K+）-ATP酶。然而，（Na++ K+）-ATP酶在刷状缘膜的比活性也增加相对于在粗质膜部分，这表明（Na++ K+）-ATP酶可能是一个内在的组成部分，除了是普遍存在于基底侧膜的肾刷状缘膜。腺苷酸环化酶与（Na++ K+）-ATP酶的分布模式相同，即在基底膜和刷状缘膜中有较高的比活性。腺苷酸环化酶在两种膜制备物中被甲状旁腺激素、降钙素、肾上腺素、异莲心素和5′-鸟苷酰亚胺二磷酸刺激。当激动剂组合使用时，增强是累加的。与腺苷酸环化酶的分布相反，鸟苷酸环化酶被发现在胞质溶胶和基底侧膜的最大比活性（NaN 3加Triton X-100）的10倍，刷状缘膜。ATP增强鸟苷酸环化酶活性只在基底侧膜。有人建议，鸟苷酸环化酶，除了（Na++ K+）-ATP酶，被用作酶的“标记”的肾基底侧膜。
Luminal brush border and contraluminal basal-lateral segments of the plasma membrane from the same kidney cortex were prepared. The brush border membrane preparation was enriched in trehalase and γ-glutamyltranspeptidase, whereas the basal-lateral membrane preparation was enriched in (Na++ K+)-ATPase. However, the specific activity of (Na++ K+)-ATPase in brush border membranes also increased relative to that in the crude plasma membrane fraction, suggesting that (Na++ K+)-ATPase may be an intrinsic constituent of the renal brush border membrane in addition to being prevalent in the basal-lateral membrane. Adenylate cyclase had the same distribution pattern as (Na++ K+)-ATPase, i.e. higher specific activity in basal-lateral membranes and present in brush border membranes. Adenylate cyclase in both membrane preparations was stimulated by parathyroid hormone, calcitonin, epinephrine, prostaglandins and 5′-guanylylimidodiphosphate. When the agonists were used in combination enhancements were additive. In contrast to the distribution of adenylate cyclase, guanylate cyclase was found in the cytosol and in basal-lateral membranes with a maximal specific activity (NaN3plus Triton X-100) 10-fold that in brush border membranes. ATP enhanced guanylate cyclase activity only in basal-lateral membranes. It is proposed that guanylate cyclase, in addition to (Na++ K+)-ATPase, be used as an enzyme “marker” for the renal basal-lateral membrane.