猪平滑肌中（Ca2+ + Mg2+）依赖性ATP酶的部分纯化及ATP依赖性Ca2+转运系统的重建。
Partial purification of (Ca2+ + Mg2+)-dependent ATPase from pig smooth muscle and reconstitution of an ATP-dependent Ca2+-transport system.
作者信息
Wuytack F, De Schutter G, Casteels R
出版信息
Biochem J. 1981 Aug 15;198(2):265-71. doi: 10.1042/bj1980265.
(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.
（钙镁）ATP酶[（Ca2+ + Mg2+）依赖性ATP酶]从猪胃（胃窦）平滑肌的微粒体部分中部分纯化得到。用脱氧胆酸盐使膜溶解，随后通过透析去除去污剂。纯化后的（钙镁）ATP酶的比活性（在37℃时）为157±12.1（7）nmol·min-1·mg-1蛋白质，并且它被钙调蛋白刺激至255±20.9（7）nmol·min·mg-1。（钙镁）ATP酶的这种纯化导致比活性增加约18倍，并且与微粒体部分相比，总酶活性的回收率为55%。部分纯化的（钙镁）ATP酶仍然含有一些Mg2+依赖性和（Na+ + K+）依赖性ATP酶活性，但其比活性的增加相对小于（钙镁）ATP酶。（钙镁）ATP酶与Mg2+依赖性和（Na+ + K+）依赖性ATP酶活性的比率分别从粗微粒体部分中的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，重构了能够进行ATP依赖性Ca2+转运的囊泡。
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