Preparation of the membrane-permeant biarsenicals FlAsH-EDT2 and ReAsH-EDT2 for fluorescent labeling of tetracysteine-tagged proteins

被引：88

作者：

Adams, Stephen R. ^{[1
]}

Tsien, Roger Y. ^{[1
,2
,3
]}

机构：

[1] Univ Calif San Diego, Dept Pharmacol, La Jolla, CA 92093 USA

[2] Univ Calif San Diego, Dept Chem & Biochem, La Jolla, CA 92093 USA

[3] Univ Calif San Diego, Howard Hughes Med Inst, La Jolla, CA 92093 USA

来源：

NATURE PROTOCOLS | 2008年 / 3卷 / 09期

关键词：

D O I：

10.1038/nprot.2008.144

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The membrane-permeant fluorogenic biarsenicals FlAsH-EDT2 and ReAsH-EDT2 can be prepared in good yields by a straightforward two-step procedure from the inexpensive precursor dyes fluorescein and resorufin, respectively. Handling of toxic reagents such as arsenic trichloride is minimized so the synthesis can be carried out in a typical chemistry laboratory, usually taking about 2-3 d. A wide range of other biarsenical reagents and intermediates that also bind to tetracysteine-tagged (CysCysProGlyCysCys) proteins can be prepared similarly using this general procedure.

引用

页码：1527 / 1534

页数：8

共 20 条

[1] New biarsenical Ligands and tetracysteine motifs for protein labeling in vitro and in vivo: Synthesis and biological applications [J].

Adams, SR ;

Campbell, RE ;

Gross, LA ;

Martin, BR ;

Walkup, GK ;

Yao, Y ;

Llopis, J ;

Tsien, RY .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2002, 124 (21) :6063-6076

[2] Labeling tetracysteine-tagged proteins with a SplAsH of color: A modular approach to bis-arsenical fluorophores [J].

Bhunia, Anjan K. ;

Miller, Stephen C. .

CHEMBIOCHEM, 2007, 8 (14) :1642-1645

[3] A red Cy3-based biarsenical fluorescent probe targeted to a complementary binding peptide [J].

Cao, Haishi ;

Xiong, Yijia ;

Wang, Ting ;

Chen, Baowei ;

Squier, Thomas C. ;

Mayer, M. Uljana .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2007, 129 (28) :8672-+

[4] CrAsH: a biarsenical multi-use affinity probe with low non-specific fluorescence [J].

Cao, Haishi ;

Chen, Baowei ;

Squier, Thomas C. ;

Mayer, M. Uljana .

CHEMICAL COMMUNICATIONS, 2006, (24) :2601-2603

[5] Site-specific labeling of cell surface proteins with biophysical probes using biotin ligase [J].

Chen, I ;

Howarth, M ;

Lin, WY ;

Ting, AY .

NATURE METHODS, 2005, 2 (02) :99-104

[6] Redirecting lipoic acid ligase for cell surface protein labeling with small-molecule probes [J].

Fernandez-Suarez, Marta ;

Baruah, Hemanta ;

Martinez-Hernandez, Laura ;

Xie, Kathleen T. ;

Baskin, Jeremy M. ;

Bertozzi, Carolyn R. ;

Ting, Alice Y. .

NATURE BIOTECHNOLOGY, 2007, 25 (12) :1483-1487

[7] Multicolor and electron microscopic imaging of connexin trafficking [J].

Gaietta, G ;

Deerinck, TJ ;

Adams, SR ;

Bouwer, J ;

Tour, O ;

Laird, DW ;

Sosinsky, GE ;

Tsien, RY ;

Ellisman, MH .

SCIENCE, 2002, 296 (5567) :503-507

[8] Golgi twins in late mitosis revealed by genetically encoded tags for live cell imaging and correlated electron microscopy [J].

Gaietta, Guido M. ;

Giepmans, Ben N. G. ;

Deerinck, Thomas J. ;

Smith, W. Bryan ;

Ngan, Lucy ;

Llopis, Juan ;

Adams, Stephen R. ;

Tsien, Roger Y. ;

Ellisman, Mark H. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (47) :17777-17782

[9] Review - The fluorescent toolbox for assessing protein location and function [J].

Giepmans, BNG ;

Adams, SR ;

Ellisman, MH ;

Tsien, RY .

SCIENCE, 2006, 312 (5771) :217-224

[10]

Griffin BA, 2000, METHOD ENZYMOL, V327, P565

← 1 2 →