pH-dependent association of carbonic anhydrase (CA) with gastric light microsomal membranes isolated from bovine abomasum. Partial characterization of membrane-associated activity.

A A López Mañanes, G R Daleo, F V Vega
Author Information
  1. A A López Mañanes: Departamento de Biologia, Facultad de Ciencias Exactas y Naturales, UNMDP, Mar del Plata, Argentina.

Abstract

1. The effect of pH on the association of carbonic anhydrase (CA) with bovine gastric light microsomal membranes (LMMs) was investigated (a) by washing LMMs containing CA activity with solutions of different pHs; (b) by studying the adsorption at various pHs of soluble bovine erythrocyte CA to washed gastric LMMs. In both cases, the association of CA with gastric LMMs was dependent on pH, being lower at neutral or alkaline pH. 2. The amount of soluble CA associated with gastric LMMs at pHs 8.0 and 9.0 was reduced when 140 mM K+/10 mM Na+ was added to the incubation medium. 3. Two sources of CA activity in bovine gastric LMMs were assumed: a loosely- and a firmly-membrane-associated activity. Both CA activities were dose-dependently inhibited by acetazolamide (I50: 3.6 x 10(-9) and 8.4 x 10(-9) M, respectively) and by chloride, acetate, iodide, bromide and nitrate at 100 mM. Firmly-membrane-associated activity appeared to be less sensitive to inhibition by acetazolamide, chloride and iodide. 4. Both activities exhibited different behavior and stability following treatment with alkaline Triton X-100. 5. The possible importance of a membrane-associated CA activity in gastric LMMs related to gastric acid secretion is discussed.

MeSH Term

Abomasum
Acetazolamide
Animals
Carbonic Anhydrase Inhibitors
Carbonic Anhydrases
Cattle
Gastric Mucosa
Hydrogen-Ion Concentration
Intracellular Membranes
Male
Microsomes
Octoxynol
Polyethylene Glycols
Potassium
Sodium

Chemicals

Carbonic Anhydrase Inhibitors
Polyethylene Glycols
Octoxynol
Sodium
Carbonic Anhydrases
Acetazolamide
Potassium

Word Cloud

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