| Abstract
| - Four-base codon strategy was applied to incorporate a fluorophore−quencher pair into specificpositions on a single protein; β-anthraniloyl-l-α,β-diaminopropionic acid (atnDap) was employed as afluorophore and p-nitrophenylalanine (ntrPhe) as a quencher. Their positions were directed by the CGGG/CCCG and GGGC/CCCG four-base codon/anticodon pairs and two doubly mutated streptavidins, i.e.,(52atnDap, 84ntrPhe) and (54ntrPhe, 84atnDap) mutants were synthesized through Escherichia coli in vitroprotein synthesizing systems. Intramolecular photoinduced electron transfer (ET) was observed as thedecrease of intensity in steady-state fluorescence spectroscopy and as the shortening of fluorescencedecaytimes. The quenching data indicated that the ET rate reflects the detailed structure of the protein.
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