T4 RNA连接酶中腺苷酸化位点的位置。

Location of the adenylylation site in T4 RNA ligase.

作者信息

Thøgersen H C, Morris H R, Rand K N, Gait M J

出版信息

Eur J Biochem. 1985 Mar 1;147(2):325-9. doi: 10.1111/j.1432-1033.1985.tb08753.x.

DOI:10.1111/j.1432-1033.1985.tb08753.x

Abstract

The purification of the enzyme T4 RNA ligase is described from an Escherichia coli strain, KR54, in which the RNA ligase gene (g63) has been inserted into the plasmid pDR540 for inducible expression of g63 from the tac promoter. Adenylylation of the purified enzyme with [14C]rATP followed by digestion with chymotrypsin yielded an adenylylated peptide, the identity of which was determined by fast-atom-bombardment mass spectrometric analysis. The results show that the AMP residue is bound covalently to the lysine at position 99 of the RNA ligase protein sequence.

摘要

描述了从大肠杆菌菌株KR54中纯化T4 RNA连接酶的方法，在该菌株中，RNA连接酶基因（g63）已插入质粒pDR540，以便从tac启动子诱导g63表达。用[14C]rATP对纯化的酶进行腺苷酸化，然后用胰凝乳蛋白酶消化，得到一个腺苷酸化肽，其身份通过快原子轰击质谱分析确定。结果表明，AMP残基共价结合到RNA连接酶蛋白质序列第99位的赖氨酸上。