DNA cloning, characterization, and inhibition studies of an α-carbonic anhydrase from the pathogenic bacterium Vibrio cholerae

J Med Chem. 2012 Dec 13;55(23):10742-8. doi: 10.1021/jm301611m. Epub 2012 Dec 4.

Abstract

We have cloned, purified, and characterized an α-carbonic anhydrase (CA, EC 4.2.1.1) from the human pathogenic bacterium Vibrio cholerae, VchCA. The new enzyme has significant catalytic activity, and an inhibition study with sulfonamides and sulfamates led to the detection of a large number of low nanomolar inhibitors, among which are methazolamide, acetazolamide, ethoxzolamide, dorzolamide, brinzolamide, benzolamide, and indisulam (KI values in the range 0.69-8.1 nM). As bicarbonate is a virulence factor of this bacterium and since ethoxzolamide was shown to inhibit the in vivo virulence, we propose that VchCA may be a target for antibiotic development, exploiting a mechanism of action rarely considered until now.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Biocatalysis
  • Carbonic Anhydrase Inhibitors / pharmacology
  • Carbonic Anhydrases / chemistry
  • Carbonic Anhydrases / drug effects
  • Carbonic Anhydrases / genetics*
  • Carbonic Anhydrases / metabolism
  • Cloning, Molecular
  • DNA / genetics*
  • Electrophoresis, Polyacrylamide Gel
  • Molecular Sequence Data
  • Phylogeny
  • Sequence Homology, Amino Acid
  • Vibrio cholerae / enzymology*

Substances

  • Carbonic Anhydrase Inhibitors
  • DNA
  • Carbonic Anhydrases