Site-specific biosynthetic incorporation of a fluorescent tag into proteins via Cysteine-tRNACys

被引：4

作者：

Lien, L ^{[1
]}

Ananda, P ^{[1
]}

Seneviratne, K ^{[1
]}

Jaikaran, ASI ^{[1
]}

Woolley, GA ^{[1
]}

机构：

[1] Univ Toronto, Dept Chem, Toronto, ON M5S 3H6, Canada

来源：

ANALYTICAL BIOCHEMISTRY | 2002年 / 307卷 / 02期

基金：

加拿大健康研究院;

关键词：

fluorescence; tRNA; cysteine; probe; protein; labeling; modification;

D O I：

10.1016/S0003-2697(02)00035-0

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Site-specific incorporation of biophysical probes into proteins during translation can permit structure/function studies on selected proteins in heterogeneous environments. We report here a procedure for incorporating a fluorescent tag into proteins via Escherichia coli Cys-tRNA(Cys) during in vitro protein synthesis. Naturally occurring Cys-tRNA(Cys) is an attractive vehicle for fluorophore incorporation since it can be readily prepared in quantity and reacted with commercially available fluorophores. Moreover, proteins can often be constructed with a single Cys so that fluorophore incorporation results in a tag at a unique site. (C) 2002 Elsevier Science (USA). All rights reserved.

引用

页码：252 / 257

页数：6

共 38 条

[1] SITE-SPECIFIC INCORPORATION OF NONNATURAL RESIDUES DURING INVITRO PROTEIN-BIOSYNTHESIS WITH SEMISYNTHETIC AMINOACYL-TRANSFER RNAS [J].