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痘苗病毒鸟苷酸转移酶对RNA的加帽作用。酶-鸟苷酸中间体的结构。

RNA capping by the vaccinia virus guanylyltransferase. Structure of enzyme-guanylate intermediate.

作者信息

Roth M J, Hurwitz J

出版信息

J Biol Chem. 1984 Nov 10;259(21):13488-94.

PMID:6092377
Abstract

GTP

RNA guanylyltransferase isolated from vaccinia virus catalyzes the transfer of GMP from GTP to the 5' terminus of RNA via an enzyme-guanylate intermediate. Incubation of the purified vaccinia RNA guanylyltransferase with [alpha- 32P]GTP and MgCl2 yields [32P]GMP covalently linked to the Mr = 95,000 subunit. The bond involves the phosphate moiety of GMP and the Ne-amino group of lysine. This was verified by treatment of the isolated 95-kDa subunit-[32P]GMP complex with sodium periodate, followed by methylamine-catalyzed beta-elimination. The product was then hydrolyzed by alkali producing 32P-labeled lysine (Ne-P)phosphate.

摘要

从牛痘病毒中分离出的GTP：RNA鸟苷酸转移酶通过酶-鸟苷酸中间体催化GMP从GTP转移至RNA的5'末端。将纯化的牛痘RNA鸟苷酸转移酶与[α-32P]GTP和MgCl2一起温育，会产生与分子量为95,000的亚基共价连接的[32P]GMP。该键涉及GMP的磷酸部分和赖氨酸的Nε-氨基。用高碘酸钠处理分离出的95-kDa亚基-[32P]GMP复合物，随后进行甲胺催化的β-消除反应，证实了这一点。然后产物用碱水解，产生32P标记的赖氨酸（Nε-P）磷酸盐。

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