title: Biochemical, histological, and inhibitor studies of membrane carbonic anhydrase in frog gastric acid secretion.
abstract: Gastric acid secretion is dependent on carbonic anhydrase (CA). To define the role of membrane-bound CA, we used biochemical, histochemical, and pharmacological approaches in the frog (Rana pipiens). CA activity and inhibition by membrane-permeant and -impermeant agents were studied in stomach homogenates and microsomal fractions. H(+) secretion in the histamine-stimulated isolated mucosa was measured before and after mucosal addition of a permeant CA inhibitor (methazolamide) and before and after mucosal or serosal addition of two impermeant CA inhibitors of differing molecular mass: a 3,500-kDa polymer linked to aminobenzolamide and p-fluorobenzyl-aminobenzolamide (molecular mass, 454 kDa). Total CA activity of frog gastric mucosa is 2,280 U/g, of which 10% is due to membrane-bound CA. Membrane-bound CA retains detectable activity below pH 4. Histochemically, there is membrane-associated CA in surface epithelial, oxynticopeptic, and capillary endothelial cells. Methazolamide reduced H(+) secretion by 100%, whereas the two impermeant inhibitors equally blocked secretion by 40% when applied to the mucosal side and by 55% when applied to the serosal side. The presence of membrane-bound CA in frog oxynticopeptic cells and its relative resistance to acid inactivation and inhibition by impermeant inhibitors demonstrate that it subserves acid secretion at both the apical and basolateral sides.
authors.affiliation: Department of Medicine, Department of Veterans Affairs Medical Center, University of Washington, Seattle, Washington 98108, USA. eswenson@u.washington.edu
authors.forename: E R
authors.initials: ER
authors.lastname: Swenson
authors.forename: T W
authors.initials: TW
authors.lastname: Tewson
authors.forename: P J
authors.initials: PJ
authors.lastname: Wistrand
authors.forename: Y
authors.initials: Y
authors.lastname: Ridderstrale
authors.forename: C
authors.initials: C
authors.lastname: Tu
mesh_terms: D010129:4-Aminobenzoic Acid / Q000494:pharmacology; D000818:Animals; D002257:Carbonic Anhydrase Inhibitors / Q000494:pharmacology; D002256:Carbonic Anhydrases / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D003432:Cross-Linking Reagents; D003593:Cytoplasm / Q000201:enzymology; D005744:Gastric Acid / Q000201:enzymology* / Q000378:metabolism; D005753:Gastric Mucosa / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D006651:Histocytochemistry; D006863:Hydrogen-Ion Concentration; D008704:Methazolamide / Q000494:pharmacology; D008861:Microsomes / Q000201:enzymology; D011092:Polyethylene Glycols / Q000494:pharmacology; D011894:Rana pipiens; D013830:Thiadiazoles / Q000494:pharmacology; D062366:para-Aminobenzoates; D005464:Fluorobenzenes; D005466:Fluorocarbons
chemical_list: D010129:4-Aminobenzoic Acid; D002257:Carbonic Anhydrase Inhibitors; D002256:Carbonic Anhydrases; D003432:Cross-Linking Reagents; D008704:Methazolamide; D011092:Polyethylene Glycols; D013830:Thiadiazoles; D062366:para-Aminobenzoates; C103638:F 3500; C068258:pentafluorobenzyl-4-aminobenzoic acid; D005464:Fluorobenzenes; D005466:Fluorocarbons