Ouabain-sensitive K(+)-ATPase in epithelial cells from guinea pig distal colon.
We have previously shown that an ouabain-sensitive H(+)-K+ exchange mechanism may be present in the apical membrane of guinea pig distal colon [Y. Suzuki and K. Kaneko. Am. J. Physiol. 256 (Gastrointest. Liver Physiol. 19): G979-G988, 1989]. The present study is aimed to demonstrate the presence of an ATPase responsible for this exchange. ATPase activity was determined in the crude membrane fraction of the colonic epithelial cell homogenate. ATPase activity under Na(+)-free conditions was increased by the addition of K+, with a half-maximal effect at 55 microM. This increase was completely abolished by 1 mM ouabain, suggesting the presence of an ouabain-sensitive K(+)-ATPase. The ouabain-sensitive K(+)-ATPase activity was inhibited by vanadate (100 microM) and N,N'-dicyclohexylcarbodiimide (100 microM) but was resistant to oligomycin (4.5 micrograms/ml) and NaN3 (1 mM). The ouabain-sensitive K(+)-ATPase activity was observed in the distal but not in the proximal colon, whereas Na(+)-K(+)-ATPase activity was distributed along the entire colon. Omeprazole (40 microM) reduced the colonic K(+)-ATPase activity by 31 +/- 6%, whereas it reduced the gastric K(+)-ATPase activity by 78 +/- 8%. These results suggest that the ouabain-sensitive K(+)-ATPase as demonstrated here is responsible for the colonic H(+)-K+ exchange. This ATPase could be similar to but is not identical with either Na(+)-K(+)-ATPase or gastric H(+)-K(+)-ATPase.
Watanabe
T
T
Department of Physiology, Yamagata University School of Medicine, Japan.
Suzuki
T
T
Suzuki
Y
Y
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D027682:Cation Transport Proteins; D002414:Cations, Monovalent; D002462:Cell Membrane / Q000201:enzymology; D003106:Colon / Q000201:enzymology*; D006224:Cricetinae; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D004848:Epithelium / Q000201:enzymology; D005753:Gastric Mucosa / Q000201:enzymology; D017506:H(+)-K(+)-Exchanging ATPase; D006863:Hydrogen-Ion Concentration; D007413:Intestinal Mucosa / Q000201:enzymology*; D007700:Kinetics; D008297:Male; D009840:Oligomycins / Q000494:pharmacology; D010042:Ouabain / Q000494:pharmacology*; D011188:Potassium / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase; D014638:Vanadates / Q000494:pharmacology
D027682:Cation Transport Proteins; D002414:Cations, Monovalent; D009840:Oligomycins; D014638:Vanadates; D004024:Dicyclohexylcarbodiimide; D010042:Ouabain; D000251:Adenosine Triphosphatases; C022761:potassium transporting ATPase; D017506:H(+)-K(+)-Exchanging ATPase; D000254:Sodium-Potassium-Exchanging ATPase; D011188:Potassium