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

FEBS Letters

Sisido, Masahiko²  Taki, Masumi²  Taira, Kazunari¹  Hohsaka, Takahiro²  Murakami, Hiroshi² 

[1] Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Hongo, Tokyo 113-8656, Japan;Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan

关键词: Non-natural mutagenesis;    Fluorescent amino acid;    In vitro protein synthesis;    Streptavidin;   

DOI  :  10.1016/S0014-5793(01)02935-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 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.

【 授权许可】

Unknown