A General and Efficient Method for the Site-Specific Dual-Labeling of Proteins for Single Molecule Fluorescence Resonance Energy Transfer

被引：146

作者：

Brustad, Eric M. ^{[1
,2
]}

Lemke, Edward A. ^{[1
,2
,3
]}

Schultz, Peter G. ^{[1
,2
]}

Deniz, Ashok A. ^{[3
]}

机构：

[1] Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA

[2] Scripps Res Inst, Skaggs Inst Chem Biol, La Jolla, CA 92037 USA

[3] Scripps Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2008年 / 130卷 / 52期

关键词：

D O I：

10.1021/ja807430h

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

A general strategy for the site-specific dual-labeling of proteins for single-molecule fluorescence resonance energy transfer is presented, A genetically encoded unnatural ketone amino acid was labeled with a hydroxylamine-containing fluorophore with high yield (>95%) and specificity. This methodology was used to construct dual-labeled T4 lysozyme variants, allowing the study of T4 lysozyme folding at single-molecule resolution. The presented strategy is anticipated to expand the scope of single-molecule protein structure and function studies.

引用

页码：17664 / +

页数：3

共 16 条

[11] Protein folding studied by single-molecule FRET [J].

Schuler, Benjamin ;

Eaton, William A. .

CURRENT OPINION IN STRUCTURAL BIOLOGY, 2008, 18 (01) :16-26

[12] Coil-globule transition in the denatured state of a small protein [J].

Sherman, Eilon ;

Haran, Gilad .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (31) :11539-11543

[13] Glycine as a D-amino acid surrogate in the K+-selectivity filter [J].

Valiyaveetil, FI ;

Sekedat, M ;

MacKinnon, R ;

Muir, TW .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (49) :17045-17049

[14] Addition of the keto functional group to the genetic code of Escherichia coli [J].

Wang, L ;

Zhang, ZW ;

Brock, A ;

Schultz, PG .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (01) :56-61

[15] Expanding the genetic code [J].

Wang, Lei ;

Xie, Jianming ;

Schultz, Peter G. .

ANNUAL REVIEW OF BIOPHYSICS AND BIOMOLECULAR STRUCTURE, 2006, 35 :225-249

[16] STRUCTURE OF BACTERIOPHAGE-T4 LYSOZYME REFINED AT 1.7 A RESOLUTION [J].

WEAVER, LH ;

MATTHEWS, BW .

JOURNAL OF MOLECULAR BIOLOGY, 1987, 193 (01) :189-199

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