Partial purification of (Ca2+ + Mg2+)-dependent ATPase from pig smooth muscle and reconstitution of an ATP-dependent Ca2+-transport system.
Partial purification of (Ca2+ + Mg2+)-dependent ATPase from pig smooth muscle and reconstitution of an ATP-dependent Ca2+-transport system.
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从猪平滑肌中部分纯化 (Ca2 Mg2) 依赖性 ATP 酶并重建 ATP 依赖性 Ca2 转运系统。
DOI：
10.1042/bj1980265
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发表时间：
1981
期刊：
影响因子：
--
通讯作者：
R. Casteels
中科院分区：
文献类型：
--
作者：
F. Wuytack;G. De Schutter;R. Casteels
（CaMg）ATP酶[（Ca ~（2+）+Mg ~（2+））-依赖性ATP酶]部分纯化自猪胃（胃窦）平滑肌的微粒体组分。用脱氧胆酸盐溶解膜，然后通过透析除去去污剂。纯化的（CaMg）ATP酶具有157 +/- 12.1（7）nmol.min-1.mg-1蛋白质的比活性（在37 ℃），并且其被钙调蛋白刺激至255 +/- 20.9（7）nmol.min-1.mg-1。（CaMg）ATP酶的这种纯化导致比活性增加约10%。18-倍，并在55%的总酶活性的回收率相比，微粒体部分。部分纯化的（CaMg）ATP酶仍具有一定的Mg ~（2+）和（Na ~+ + K ~+）依赖的ATP酶活性，但其比活性的提高幅度相对较小。（CaMg）ATP酶与Mg ~（2+）和（Na ~+ + K ~+）依赖的ATP酶活性的比值分别从粗微粒体组分中的0.14和0.81增加到纯化制剂中的1.39和9.07。在通过透析去除脱氧胆酸盐的过程中，囊泡被重构，其能够进行ATP依赖性Ca 2+转运。
(CaMg)ATPase [(Ca2+ + Mg2+)-dependent ATPase] was partially purified from a microsomal fraction of the smooth muscle of the pig stomach (antrum). Membranes were solubilized with deoxycholate, followed by removal of the detergent by dialysis. The purified (CaMg)ATPase has a specific activity (at 37 degrees C) of 157 +/- 12.1 (7)nmol.min-1.mg-1 of protein, and it is stimulated by calmodulin to 255 +/- 20.9 (7)nmol.min.mg-1. This purification of the (CaMg)ATPase resulted in an increase of the specific activity by approx. 18-fold and in a recovery of the total enzyme activity of 55% compared with the microsomal fraction. The partially purified (CaMg)ATPase still contains some Mg2+-and (Na+ + K+)-dependent ATPase activities, but their specific activities are increased relatively less than that of the (CaMg)ATPase. The ratios of the (CaMg)ATPase to Mg2+- and (Na+ + K+)-dependent ATPase activities increase from respectively 0.14 and 0.81 in the crude microsomal fraction to 1.39 and 9.07 in the purified preparation. During removal of the deoxycholate by dialysis, vesicles were reconstituted which were capable of ATP-dependent Ca2+ transport.