活性α亚基从二聚体αβ禽成髓细胞瘤病毒DNA聚合酶释放的机制。
Mechanism of release of active alpha subunit from dimeric alpha beta avian myeloblastosis virus DNA polymerase.
作者信息
Papas T S, Marciani D J, Samuel K, Chirikjian J G
出版信息
J Virol. 1976 Jun;18(3):904-10. doi: 10.1128/JVI.18.3.904-910.1976.
Abstract
Storage of the dimeric (alphabeta) form of avian myeloblastosis virus (AMV) DNA polymerase in glycerol resulted in the release of the smaller alpha subunit, as detected by glycerol gradient sedimentation. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of enzyme stored in glycerol showed the concomitant appearance of several polypeptides and a lowering in the level of both beta and alpha components. This reduction appears to be the result of cleavages introduced by traces of hydrolytic activity present in glycerol samples. An enhancement of alpha subunit released, as detected by activity profile, was also achieved upon direct but limited exposure of purified avian myeloblastosis virus DNA polymerase to carboxymethyl-cellulose-bound trypsin matrix. Electrophoretic analysis of digested enzyme revealed a progressive fragmentation, with simultaneous increase in the alpha subunit and decrease in the beta subunit.
摘要
通过甘油梯度沉降检测发现，禽成髓细胞瘤病毒（AMV）DNA聚合酶的二聚体（αβ）形式储存在甘油中会导致较小的α亚基释放出来。对储存在甘油中的酶进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析显示，同时出现了几种多肽，并且β和α组分的水平降低。这种降低似乎是甘油样品中存在的微量水解活性所导致的裂解结果。通过活性分析检测到，将纯化的禽成髓细胞瘤病毒DNA聚合酶直接但有限地暴露于羧甲基纤维素结合的胰蛋白酶基质中，也能提高α亚基的释放量。对消化后的酶进行电泳分析发现，酶会逐渐碎片化，同时α亚基增加而β亚基减少。
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