AZIDE-SPECIFIC LABELING OF BIOMOLECULES BY STAUDINGER-BERTOZZI LIGATION: PHOSPHINE DERIVATIVES OF FLUORESCENT PROBES SUITABLE FOR SINGLE-MOLECULE FLUORESCENCE SPECTROSCOPY

被引：17

作者：

Chakraborty, Anirban ^{[1
,2
]}

Wang, Dongye ^{[1
,2
]}

Ebright, Yon W. ^{[1
,2
]}

Ebright, Richard H. ^{[1
,2
]}

机构：

[1] Rutgers State Univ, Waksman Inst, Dept Chem & Biol Chem, Piscataway, NJ 08854 USA

[2] Rutgers State Univ, Howard Hughes Med Inst, Piscataway, NJ USA

来源：

METHODS IN ENZYMOLOGY, VOL 472: SINGLE MOLECULE TOOLS, PT A: FLUORESCENCE BASED APPROACHES | 2010年 / 472卷

关键词：

PHOTO-CROSS-LINKING; RECOMBINANT PROTEINS; ESCHERICHIA-COLI; LIVING ANIMALS; GENETIC-CODE; AMINO-ACIDS; GLYCANS; VISUALIZATION; FUNCTIONALITY; CHEMISTRY;

D O I：

10.1016/S0076-6879(10)72018-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe the synthesis of phosphine derivatives of three fluorescent probes that have a brightness and photostability suitable for single-molecule fluorescence spectroscopy and microscopy: Alexa488, Cy3B, and Alexa647. In addition, we describe procedures for use of these reagents in azide-specific, bioorthogonal labeling through Staudinger-Bertozzi ligation, as well as procedures for the quantitation of labeling specificity and labeling efficiency. The reagents and procedures of this report enable chemoselective, site-selective labeling of azide-containing biomolecules for single-molecule fluorescence spectroscopy and microscopy.

引用

页码：19 / 30

页数：12

共 33 条

[1] An engineered aryl azide ligase for site-specific mapping of protein-protein interactions through photo-cross-linking [J].