来自雪白根霉的碱基特异性核糖核酸酶Ru的纯化

Purification of a base-specific ribonuclease Ru from Rhizopus niveus.

作者信息

Horitsu H, Takihi N, Sugiura M, Tomoyeda M

出版信息

Can J Biochem. 1980 Jun;58(6):489-93. doi: 10.1139/o80-065.

Abstract

A base-specific ribonuclease (RNase) Ru (EC 3.1.27.5) was isolated and purified from Rhizopus niveus in a yield of 17% by the procedures of acetone precipitation, column chromatography on Duolite A-2, DEAE-cellulose, CM-cellulose, and 2'(3')-aminohexyl-5'-UMP-agarose. The enzyme was shown to be homogeneous by polyacrylamide disc electrophoresis. The amino- and carboxyl-terminal amino acids of the enzyme were determined to be an arginine and an aspartic acid, respectively. The enzyme has a base specificity: it released only 3'-UMP from yeast RNA or poly(U) and, in addition, small amounts of 3'-CMP from poly(C).

摘要

通过丙酮沉淀、在Duolite A - 2、DEAE - 纤维素、CM - 纤维素和2'(3') - 氨基己基 - 5'-UMP - 琼脂糖上进行柱色谱等步骤，从雪白根霉中分离并纯化出了一种碱基特异性核糖核酸酶（RNase）Ru（EC 3.1.27.5），产率为17%。通过聚丙烯酰胺圆盘电泳表明该酶是纯的。该酶的氨基末端和羧基末端氨基酸分别被确定为精氨酸和天冬氨酸。该酶具有碱基特异性：它仅从酵母RNA或聚（U）中释放3'-UMP，此外，还从聚（C）中释放少量的3'-CMP。