Characterization, Bioinformatic Analysis and Dithiocarbamate Inhibition Studies of Two New α-carbonic Anhydrases, CAH1 and CAH2, From the Fruit Fly Drosophila Melanogaster

Bioorg Med Chem. 2013 Mar 15;21(6):1516-21. doi: 10.1016/j.bmc.2012.08.046. Epub 2012 Aug 31.

Abstract

Carbonic anhydrases (CAs) are essential and ubiquitous enzymes. Thus far, there are no articles on characterization of Drosophila melanogaster α-CAs. Data from invertebrate CA studies may provide opportunities for anti-parasitic drug development because α-CAs are found in many parasite or parasite vector invertebrates. We have expressed and purified D. melanogaster CAH1 and CAH2 as proteins of molecular weights 30kDa and 28kDa. CAH1 is cytoplasmic whereas CAH2 is a membrane-attached protein. Both are highly active enzymes for the CO2 hydration reaction, being efficiently inhibited by acetazolamide. CAH2 in the eye of D. melanogaster may provide a new animal model for CA-related eye diseases. A series of dithiocarbamates were also screened as inhibitors of these enzymes, with some representatives showing inhibition in the low nanomolar range.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbonic Anhydrase I / antagonists & inhibitors*
  • Carbonic Anhydrase I / classification
  • Carbonic Anhydrase I / metabolism
  • Carbonic Anhydrase II / antagonists & inhibitors*
  • Carbonic Anhydrase II / classification
  • Carbonic Anhydrase II / metabolism
  • Carbonic Anhydrase Inhibitors / chemistry*
  • Carbonic Anhydrase Inhibitors / metabolism
  • Computational Biology
  • Drosophila melanogaster / enzymology*
  • Kinetics
  • Phylogeny
  • Protein Binding
  • Recombinant Fusion Proteins / antagonists & inhibitors
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / isolation & purification
  • Thiocarbamates / chemistry*
  • Thiocarbamates / metabolism

Substances

  • Carbonic Anhydrase Inhibitors
  • Recombinant Fusion Proteins
  • Thiocarbamates
  • Carbonic Anhydrase I
  • Carbonic Anhydrase II