Limited proteolysis of the erythrocyte membrane skeleton by calcium-dependent proteinases.
Limited proteolysis of the erythrocyte membrane skeleton by calcium-dependent proteinases.
复制标题
钙依赖性蛋白酶对红细胞膜骨架的有限蛋白水解。
DOI：
10.1016/0304-4165(86)90250-3
复制
发表时间：
1986
期刊：
影响因子：
--
通讯作者：
DeMartino,GN
中科院分区：
文献类型：
--
作者：
Croall,DE;Morrow,JS;DeMartino,GN
本文研究了纯化的钙依赖蛋白酶对人红细胞膜骨架蛋白的作用。当钙依赖性蛋白酶I或钙依赖性蛋白酶II在钙存在下进入血影膜骨架的胞质侧时，发生蛋白4.1 a和B以及条带3的优先切割以及α-和β-血影蛋白的有限切割。因此，当这些蛋白酶与密封的鬼孵育时，它们不切割这些蛋白质。亮抑酶肽、mersalyl（这些酶的特异性细胞蛋白抑制剂）和钙螯合剂可抑制Ca 2+依赖性蛋白酶对红细胞血影蛋白的蛋白水解。每种蛋白酶也在低离子强度下在不存在钙的情况下被加载到红细胞血影中，随后通过重新密封血影而被困在其中。通过在钙离子载体A23187和钙的存在下孵育这些血影来激活蛋白酶。通过电泳对血影蛋白的检查表明，条带4.1和3的钙依赖性蛋白水解以及α-和β-血影蛋白的有限切割与开放、渗漏血影蛋白的蛋白水解观察到的结果相似。在钙存在的情况下，每种钙依赖性蛋白酶似乎与红细胞血影膜有关。
The action of purified calcium-dependent proteinase on human erythrocyte membrane skeleton proteins has been examined. Preferential cleavage of proteins 4.1 a and b and band 3 and limited cleavage of α- and β-spectrin occur when either calcium-dependent proteinase I or calcium-dependent proteinase II has access to the cytoplasmic side of the ghost membrane skeleton in the presence of calcium. Thus, when these proteinases are incubated with sealed ghosts they do not cleave these proteins. Leupeptin, mersalyl, the specific cellular protein inhibitor of these enzymes, and calcium chelators can inhibit proteolysis of the red cell ghost proteins by Ca2+-dependent proteinases. Each proteinase has also been loaded into erythrocyte ghosts in the absence of calcium at low ionic strength and subsequently trapped inside by resealing the ghosts. The proteinases were activated by incubating these ghosts in the presence of the calcium ionophore A23187 and calcium. Examination of the ghost proteins by electrophoresis demonstrated calcium-dependent proteolysis of Bands 4.1 and 3 and limited cleavage of α- and β-spectrin similar to that observed on proteolysis of the open, leaky ghosts. In the presence of calcium each calcium-dependent proteinase appears to associate with the erythrocyte ghost membrane.