Multicolor protein FRET with tryptophan, selective coumarin-cysteine labeling, and genetic acridonylalanine encoding

被引：21

作者：

Ferrie, John J. ^{[1
]}

Ieda, Naoya ^{[1
,2
]}

Haney, Conor M. ^{[1
]}

Walters, Christopher R. ^{[1
]}

Sungwienwong, Itthipol ^{[1
]}

Yoon, Jimin ^{[1
]}

Petersson, E. James ^{[1
]}

机构：

[1] Univ Penn, Dept Chem, 213 South 34th St, Philadelphia, PA 19104 USA

[2] Nagoya City Univ, Grad Sch Pharmaceut Sci, Dept Organ & Med Chem, Mizuho Ku, 3-1 Tanabedori, Nagoya, Aichi 4678603, Japan

来源：

CHEMICAL COMMUNICATIONS | 2017年 / 53卷 / 80期

基金：

美国国家卫生研究院; 日本学术振兴会;

关键词：

RESONANCE ENERGY-TRANSFER; AMINO-ACID; ALPHA-SYNUCLEIN; PEPTIDE COMPLEXES; CALMODULIN; DISSOCIATION; LANDSCAPE; STABILITY; CHANNEL; CELLS;

D O I：

10.1039/c7cc05492k

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Site-specific fluorescence probes can be used to measure distances within proteins when used as part of a Forster resonance energy transfer (FRET) pair. Here we report the synthesis of a coumarin maleimide (Mcm-Mal) that is fluorogenic upon reaction with cysteine. We demonstrate that cysteine, acridonylalanine (Acd) double mutant proteins can be produced by unnatural amino acid mutagenesis and reacted with Mcm-Mal to generate Mcm/Acd labeled proteins for FRET studies. The Mcm/Acd FRET pair is minimally-perturbing, easy to install, and well-suited to studying protein distances in the 15-40 angstrom range. Furthermore, Mcm/Acd labeling can be combined with tryptophan fluorescence in three color FRET to monitor multiple interactions in one experiment.

引用

页码：11072 / 11075

页数：4

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