Article
Resolution of two substrate-binding sites in an engineered cytochrome P450eryF bearing a fluorescent probe.
Biophysical journal - 1 Jul 2005
Davydov Dmitri R, Botchkareva Alexandra E, Davydova Nadezhda E, Halpert James R
Abstract excerpt
To elucidate the mechanisms of cooperativity of cytochrome P450eryF an SH-reactive fluorescent probe was introduced close to the substrate-binding site. Cys-154, the only accessible cysteine, was eliminated by site-directed mutagenesis, and a novel cysteine was substituted for Ser-93 in the B'/C loop. S93C, C154A, C154S, S93C/C154A, and S93C/S154C were characterized in terms of affinity for 1-pyrenebutanol...
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