HatoTag: A novel protein labeling technology for cell imaging and protein analysis

被引：1641

作者：

Los, Georgyi V. ^{[1
]}

Encell, Lance P. ^{[1
]}

McDougall, Mark G. ^{[2
]}

Hartzell, Danette D. ^{[1
]}

Karassina, Natasha ^{[1
]}

Zimprich, Chad ^{[1
]}

Wood, Monika G. ^{[1
]}

Learish, Randy ^{[1
]}

Ohane, Rachel Friedman ^{[1
]}

Urh, Marjeta ^{[1
]}

Simpson, Dan ^{[1
]}

Mendez, Jacqui ^{[1
]}

Zimmerman, Kris ^{[1
]}

Otto, Paul ^{[1
]}

Vidugiris, Gediminas ^{[1
]}

Zhu, Ji ^{[2
]}

Darzins, Aldis ^{[1
,3
]}

Klaubert, Dieter H. ^{[2
]}

Bulleit, Robert F. ^{[1
]}

Wood, Keith V. ^{[1
]}

机构：

[1] Promega Corp, Madison, WI 53711 USA

[2] Promega Biosci Inc, San Luis Obispo, CA 93401 USA

[3] Natl Renewable Energy Lab, Golden, CO 80401 USA

来源：

ACS CHEMICAL BIOLOGY | 2008年 / 3卷 / 06期

关键词：

D O I：

10.1021/cb800025k

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：

We have designed a modular protein tagging system that allows different functionalities to be linked onto a single genetic fusion, either in solution, in living cells, or in chemically fixed cells. The protein tag (HaloTag) is a modified haloalkane dehalogenase designed to covalently bind to synthetic ligands (HaloTag ligands). The synthetic ligands comprise a chloroalkane linker attached to a variety of useful molecules, such as fluorescent dyes, affinity handles, or solid surfaces. Covalent bond formation between the protein tag and the chloroalkane linker is highly specific, occurs rapidly under physiological conditions, and is essentially irreversible. We demonstrate the utility of this system for cellular imaging and protein immobilization by analyzing multiple molecular processes associated with NF-kappa B-mediated cellular physiology, including imaging of subcellular protein translocation and capture of protein-protein and protein-DNA complexes.

引用

页码：373 / 382

页数：10

共 33 条

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