Structure and mechanism of a bacterial haloalcohol dehalogenase: a new variation of the short-chain dehydrogenase/reductase fold without an NAD(P)H binding site

R.M.de Jong, J.J.W. Tiesinga, H.J. Rozeboom, K.H. Kalk, D.B. Janssen, B.W. Dijkstra

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Abstract

Haloalcohol dehalogenases are bacterial enzymes that catalyze the cofactor-independent dehalogenation of vicinal haloalcohols such as the genotoxic environmental pollutant 1,3-dichloro-2-propanol, thereby producing an epoxide, a chloride ion and a proton. Here we present X-ray structures of the haloalcohol dehalogenase HheC from Agrobacterium radiobacter AD1, and complexes of the enzyme with an epoxide product and chloride ion, and with a bound haloalcohol substrate mimic. These structures support a catalytic mechanism in which Tyr145 of a Ser-Tyr-Arg catalytic triad deprotonates the haloalcohol hydroxyl function to generate an intramolecular nucleophile that substitutes the vicinal halogen. Haloalcohol dehalogenases are related to the widespread family of NAD(P)H-dependent short-chain dehydrogenases/reductases (SDR family), which use a similar Ser-Tyr-Lys/Arg catalytic triad to catalyze reductive or oxidative conversions of various secondary alcohols and ketones. Our results reveal the first structural details of an SDR-related enzyme that catalyzes a substitutive dehalogenation reaction rather than a redox reaction, in which a halide-binding site is found at the location of the NAD(P)H binding site. Structure-based sequence analysis reveals that the various haloalcohol dehalogenases have likely originated from at least two different NAD-binding SDR precursors.

Original languageEnglish
Pages (from-to)4933-4944
Number of pages12
JournalThe EMBO Journal
Volume22
Issue number19
DOIs
Publication statusPublished - 1-Oct-2003

Keywords

  • haloalcohol dehalogenase
  • SDR family
  • short-chain dehydrogenase
  • X-ray structure
  • XANTHOBACTER-AUTOTROPHICUS GJ10
  • AGROBACTERIUM-RADIOBACTER AD1
  • ALPHA/BETA-HYDROLASE FOLD
  • 1.9 ANGSTROM RESOLUTION
  • CRYSTAL-STRUCTURES
  • DEHYDROGENASES/REDUCTASES SDRS
  • L-2-HALOACID DEHALOGENASE
  • HALOALKANE DEHALOGENASE
  • HALOHYDRIN DEHALOGENASE
  • FUNCTIONAL ASSIGNMENTS

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