Chemical synthesis of 4-thiouridine-containing substrate analogues of tRNA:pseudouridine-55 synthase:: Photocross-linking studies

被引：3

作者：

Becker, HF

Grosjean, H

Fourrey, JL ^{[1
]}

机构：

[1] CNRS, Inst Chim Subst Nat, F-91198 Gif Sur Yvette, France

[2] CNRS, Lab Enzymol & Biochim Struct, F-75700 Paris, France

来源：

NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS | 1998年 / 17卷 / 12期

关键词：

D O I：

10.1080/07328319808004327

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

For site specific incorporation of 4-thiouridine into oligoribonucleotides a new phosphoramidite is proposed. It makes use of the S-pivaloyloxymethyl group for the protection of the thiol function. This group is easily introduced and removed without modification of the standard protocol for solid phase synthesis of RNA. Three 4-thiouridine-containing oligoribonucleotides (21-mers), corresponding to tRNA minisubstrates of yeast tRNA:pseudouridine-55 synthase (Pus4) were prepared. These 4-thiouridine containing substrates were characterized and used as photoaffinity probe of the enzyme:substrate complex. Irradiation resulted in the specific photocross-linking of these oligoribonucleotides with purified recombinant tRNA:pseudouridine-55 synthase.

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页码：2403 / 2416

页数：14

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