Tryptophan 7-halogenase (PrnA) structure suggests a mechanism for regioselective chlorination

Changjiang Dong; Silvana Flecks; Susanne Unversucht; Caroline Haupt; Karl-Heinz van Pée; James H Naismith

doi:10.1126/science.1116510

Tryptophan 7-halogenase (PrnA) structure suggests a mechanism for regioselective chlorination

Changjiang Dong, Silvana Flecks, Susanne Unversucht, Caroline Haupt, Karl-Heinz van Pée, James H Naismith

Research output: Contribution to journal › Article › peer-review

321 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	2216-9
Number of pages	4
Journal	Science
Volume	309
Issue number	5744
DOIs	https://doi.org/10.1126/science.1116510
Publication status	Published - 30 Sep 2005

Keywords

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

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10.1126/science.1116510

Cite this

@article{922a0c7451df496089be1712b043ae8d,

title = "Tryptophan 7-halogenase (PrnA) structure suggests a mechanism for regioselective chlorination",

abstract = "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.",

keywords = "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",

author = "Changjiang Dong and Silvana Flecks and Susanne Unversucht and Caroline Haupt and {van P{\'e}e}, Karl-Heinz and Naismith, {James H}",

year = "2005",

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doi = "10.1126/science.1116510",

language = "English",

volume = "309",

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TY - JOUR

T1 - Tryptophan 7-halogenase (PrnA) structure suggests a mechanism for regioselective chlorination

AU - Dong, Changjiang

AU - Flecks, Silvana

AU - Unversucht, Susanne

AU - Haupt, Caroline

AU - van Pée, Karl-Heinz

AU - Naismith, James H

PY - 2005/9/30

Y1 - 2005/9/30

N2 - 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.

AB - 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.

KW - Amino Acid Sequence

KW - Binding Sites

KW - Chlorides

KW - Crystallography, X-Ray

KW - Dimerization

KW - Flavin-Adenine Dinucleotide

KW - Hydrogen Bonding

KW - Hypochlorous Acid

KW - Indoles

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Oxidation-Reduction

KW - Oxidoreductases

KW - Oxygen

KW - Protein Conformation

KW - Protein Structure, Secondary

KW - Protein Structure, Tertiary

KW - Pseudomonas fluorescens

KW - Tryptophan

U2 - 10.1126/science.1116510

DO - 10.1126/science.1116510

M3 - Article

C2 - 16195462

VL - 309

SP - 2216

EP - 2219

JO - Science

JF - Science

SN - 0036-8075

IS - 5744

ER -