1-5-dihydro-fad and 7-chlorotryptophan

1-5-dihydro-fad has been researched along with 7-chlorotryptophan* in 2 studies

Other Studies

2 other study(ies) available for 1-5-dihydro-fad and 7-chlorotryptophan

ArticleYear
Robust in vitro activity of RebF and RebH, a two-component reductase/halogenase, generating 7-chlorotryptophan during rebeccamycin biosynthesis.
    Proceedings of the National Academy of Sciences of the United States of America, 2005, Mar-15, Volume: 102, Issue:11

    The indolocarbazole antitumor agent rebeccamycin is modified by chlorine atoms on each of two indole moieties of the aglycone scaffold. These halogens are incorporated during the initial step of its biosynthesis from conversion of L-Trp to 7-chlorotryptophan. Two genes in the biosynthetic cluster, rebF and rebH, are predicted to encode the flavin reductase and halogenase components of an FADH2-dependent halogenase, a class of enzymes involved in the biosynthesis of numerous halogenated natural products. Here, we report that, in the presence of O2, chloride ion, and L-Trp as cosubstrates, purified RebH displays robust regiospecific halogenating activity to generate 7-chlorotryptophan over at least 50 catalytic cycles. Halogenation by RebH required the addition of RebF, which catalyzes the NADH-dependent reduction of FAD to provide FADH2 for the halogenase. Maximal rates were achieved at a RebF/RebH ratio of 3:1. In air-saturated solutions, a k(cat) of 1.4 min(-1) was observed for the RebF/RebH system but increased at least 10-fold in low-pO2 conditions. RebH was also able to use bromide ions to generate monobrominated Trp. The demonstration of robust chlorinating activity by RebF/RebH sets up this system for the probing of mechanistic questions regarding this intriguing class of enzymes.

    Topics: Carbazoles; Chromatography, High Pressure Liquid; Flavin-Adenine Dinucleotide; Indoles; Kinetics; Oxidoreductases; Tryptophan

2005
Tryptophan 7-halogenase (PrnA) structure suggests a mechanism for regioselective chlorination.
    Science (New York, N.Y.), 2005, Sep-30, Volume: 309, Issue:5744

    Chlorinated natural products include vancomycin and cryptophycin A. Their biosynthesis involves regioselective chlorination by flavin-dependent halogenases. We report the structural characterization of tryptophan 7-halogenase (PrnA), which regioselectively chlorinates tryptophan. Tryptophan and flavin adenine dinucleotide (FAD) are separated by a 10 angstrom-long tunnel and bound by distinct enzyme modules. The FAD module is conserved in halogenases and is related to flavin-dependent monooxygenases. On the basis of biochemical studies, crystal structures, and by analogy with monooxygenases, we predict that FADH2 reacts with O2 to make peroxyflavin, which is decomposed by Cl-. The resulting HOCl is guided through the tunnel to tryptophan, where it is activated to participate in electrophilic aromatic substitution.

    Topics: Amino Acid Sequence; Binding Sites; Chlorides; Crystallography, X-Ray; Dimerization; Flavin-Adenine Dinucleotide; Hydrogen Bonding; Hypochlorous Acid; Indoles; Models, Molecular; Molecular Sequence Data; Oxidation-Reduction; Oxidoreductases; Oxygen; Protein Conformation; Protein Structure, Secondary; Protein Structure, Tertiary; Pseudomonas fluorescens; Tryptophan

2005