Bacterial flavodoxins are models for interaction of the FMN-containing domain of P450 reductase with microsomal P450S

C. M. Jenkins, M. R. Waterman

Research output: Contribution to journalArticlepeer-review

Abstract

Microsomal P450s expressed in f. coli are enzymatically active in intact bacteria and this activity has been found to be supported by the two component flavodoxin (FMN)/flavodoxin reductase (FAD) system. Flavodoxin binds to P450s (Ks s 1 //M) and can serve as a model to study the interaction of P450 reductase with P450s. Mutation of acidic residues in Anabaena flavodoxin which correlate with similar mutations in acidic cluster I of P450 reductase {Shen and Kasper) significantly decreases P450c17 activity while having no effect on flavodoxin binding to the P450. Apparently, even though no obvious signature sequence for reductase binding is present in microsomal P450s (unlike mitochondrial P450s), specific charge pairing interactions are required to properly orient the reductase FMN with the P450 heme to facilitate electron transfer. Efforts to locate crucial lysine residues on P450c17 by chemical modification studies plus/minus flavodoxin have not yet identified such key residues. However chemical modification has localized lysine residues associated with loss of enzymatic activity. We have used sulfo-NHS-biotin modification because it permits affinity purification of modified peptides. Based on results obtained in this and other laboratories, the nature of binding sites for P450 reductase on P450s will be discussed.

Original languageEnglish
Pages (from-to)A771
JournalFASEB Journal
Volume11
Issue number9
StatePublished - Dec 1 1997

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