Cloning, polymorphism, and inhibition of β-carbonic anhydrase of Helicobacter pylori

Carbonic anhydrase (CA) catalyzes the reversible hydration of CO2 to bicarbonate and a proton, and alpha-class CA has been reported to facilitate the acid acclimation of Helicobacter pylori (hp alpha CA). The purpose of this study was to characterize the beta-class CA of H. pylori (hp beta CA) and elucidate the role of this enzyme as a possible drug target for eradication therapy. We isolated DNA clones of independent H. pylori strains obtained from patients with gastritis (n = 15), gastric ulcer (n = 6), or gastric cancer (n = 16), and then studied genetic polymorphisms. In addition, the susceptibility of H. pylori to sulpiride, an antiulcer drug and efficient inhibitor of both hp alpha CA and hp beta CA, was studied with an in vitro killing assay. DNA sequences of all 37 hp beta CA clones encoded a 221 amino acid polypeptide with a variety of polymorphisms (57 types of amino acid substitution at 48 residue positions). There was no polymorphism functionally relevant to the gastric lesion type. One strain included unique residues that were not seen in the other 36 clones from Japanese patients but which were found in a strain obtained from the United Kingdom. Sulpiride had killing effects at concentrations greater than 200 mu g/ml for H. pylori, including strains resistant to clarithromycin, metronidazole, or ampicillin. Helicobacter pylori might have evolved independently in the Caucasian and Japanese populations. Dual inhibition of alpha-and beta-class CAs could be applied as alternative therapy for eradication of H. pylori.