title: Expression and localization of alpha- and beta-carbonic anhydrase in Helicobacter pylori.
abstract: Helicobacter pylori, the causative agent of peptic ulcer disease, expresses two different forms of the zinc-containing enzyme carbonic anhydrase (CA) (alpha and beta), catalyzing the reversible hydration of CO(2). Presumably, the high CO(2) requirement of H. pylori implies an important role for this enzyme in the bacterial physiology. In this paper, expression of the CAs has been analyzed in three different strains of the bacterium, 26695, J99 and 17.1, and appears to be independent of CO(2) concentration in the investigated range (0.1-10%). Presence of the potent and highly specific CA inhibitor, acetazolamide, in the medium does not seem to inhibit bacterial growth at the given sulfonamide concentration. Moreover, the localization and distribution of the alpha-CA was analyzed by immunonegative staining, while SDS-digested freeze-fracture immunogold labelling was used for the beta-form of the enzyme. The latter method has the advantage of allowing assessment of protein localization to distinct cell compartments and membrane structures. The resulting electron microscopy images indicate a localization of the beta-CA in the cytosol, on the cytosolic side of the inner membrane and on the outer membrane facing the periplasmic space. The alpha-enzyme was found attached to the surface of the bacterium.
authors.affiliation: Department of Chemistry, Biochemistry, Umeå University, SE-90187 Umeå, Sweden.
authors.forename: Laura C
authors.initials: LC
authors.lastname: Chirica
authors.forename: Christoffer
authors.initials: C
authors.lastname: Petersson
authors.forename: Marina
authors.initials: M
authors.lastname: Hurtig
authors.forename: Bengt-Harald
authors.initials: BH
authors.lastname: Jonsson
authors.forename: Thomas
authors.initials: T
authors.lastname: Borén
authors.forename: Sven
authors.initials: S
authors.lastname: Lindskog
mesh_terms: D002256:Carbonic Anhydrases / Q000235:genetics* / Q000378:metabolism; D005614:Freeze Fracturing; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D016680:Genome, Bacterial; D016480:Helicobacter pylori / Q000201:enzymology* / Q000235:genetics* / Q000648:ultrastructure; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism
chemical_list: D007527:Isoenzymes; D002256:Carbonic Anhydrases