A non-natural amino acid for efficient incorporation into proteins as a sensitive fluorescent probe

被引：44

作者：

Taki, M

Hohsaka, T

Murakami, H

Taira, K

Sisido, M

机构：

[1] Okayama Univ, Fac Engn, Dept Biosci & Biotechnol, Okayama 7008530, Japan

[2] Univ Tokyo, Grad Sch Engn, Dept Chem & Biotechnol, Tokyo 1138656, Japan

来源：

FEBS LETTERS | 2001年 / 507卷 / 01期

基金：

日本学术振兴会;

关键词：

non-natural mutagenesis; fluorescent amino acid; in vitro protein synthesis; streptavidin;

D O I：

10.1016/S0014-5793(01)02935-0

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：

A small and highly fluorescent non-natural amino acid that contains an anthraniloyl group (atnDap) was incorporated into various positions of streptavidin. The positions were directed by a CGGG/CCCG four-base codon/anticodon pair. The non-natural mutants were obtained in excellent yields and some of them retained strong biotin-binding activity. The fluorescence wavelength as well as the intensity of the anthraniloyl group at position 120 were sensitive to biotin binding. These unique properties indicate that the atnDap is the most suitable non-natural amino acid for a position-specific fluorescent labeling of proteins that is highly sensitive to microenvironmental changes. (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

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页码：35 / 38

页数：4

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