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Position-specific incorporation of a highly photodurable and blue-laser excitable fluorescent amino acid into proteins for fluorescence sensing.

Abstract：

:A new fluorescent amino acid, L-2-acridonylalanine, was incorporated into proteins at specific positions using 4-base codon/anticodon strategy. The efficiency of the incorporation was high enough to obtain enough quantities of the mutants. The acridonyl group was highly fluorescent when it was excited at the wavelengths of blue-lasers and was highly photodurable compared with conventional fluorophores often used for biological analyses. The fluorescence intensity was sensitive to small changes in the polarity of the environment. When the nonnatural amino acid was incorporated into specific positions of streptavidin, the mutant protein worked as a fluorescent sensor to biotin. Similarly, when the amino acid was incorporated into camel single-chain antibody, the mutant protein sensitively responded to the antigen molecule. The high incorporation efficiency, the high photodurability, the excitability with blue-lasers, and high sensitivity to the environment make the acridonylalanine as the promising fluorescent amino acid for sensing small molecules when incorporated into specific positions of various antibodies, receptors, and enzymes.

journal_name

Bioorg Med Chem

journal_title

Bioorganic & medicinal chemistry

authors

Hamada H,Kameshima N,Szymańska A,Wegner K,Lankiewicz Ł,Shinohara H,Taki M,Sisido M

doi

10.1016/j.bmc.2005.03.014

subject

Has Abstract

pub_date

2005-05-16 00:00:00

pages

3379-84

issue

10

eissn

0968-0896

issn

1464-3391

pii

S0968-0896(05)00209-9

journal_volume

13

pub_type

杂志文章

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