Site-Specific N- and C-Terminal Labeling of a Single Polypeptide Using Sortases of Different Specificity

被引：186

作者：

Antos, John M. ^{[1
]}

Chew, Guo-Liang ^{[1
]}

Guimaraes, Carla P. ^{[1
]}

Yoder, Nicholas C. ^{[1
]}

Grotenbreg, Gijsbert M. ^{[1
]}

Popp, Maximilian Wei-Lin ^{[1
]}

Ploegh, Hidde L. ^{[1
]}

机构：

[1] Whitehead Inst Biomed Res, Cambridge, MA 02142 USA

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2009年 / 131卷 / 31期

基金：

美国国家卫生研究院;

关键词：

SURFACE PROTEIN;

D O I：

10.1021/ja902681k

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

The unique reactivity of two sortase enzymes, SrtA(staph) from Staphylococcus aureus and SrtA(strep) from Streptococcus pyogenes, is exploited for site-specific labeling of a single polypeptide with different labels at its N and C termini. SrtA(strep) is used to label the protein's C terminus at an LPXTG site with a fluorescently labeled dialanine nucleophile. Selective N-terminal. labeling of proteins containing N-terminal glycine residues is achieved using SrtA(staph) and LPXT derivatives. The generality of N-terminal labeling with SrtA(staph) is demonstrated by near-quantitative labeling of multiple protein substrates with excellent site specificity.

引用

收藏

页码：10800 / +

页数：3

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