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J Biol Chem, Vol. 274, Issue 28, 19693-19698, July 9, 1999

The Colonic H⁺,K⁺-ATPase Functions as a Na⁺-dependent K⁺(NH₄⁺)-ATPase in Apical Membranes from Rat Distal Colon

Juan Codina, Thomas A. Pressley^§, and Thomas D. DuBose Jr.

From the  Division of Renal Diseases and Hypertension Department of Internal Medicine, University of Texas, Houston Medical School, Houston, Texas 77030 and the ^§  Department of Physiology, Texas Tech University Health Sciences Center, Lubbock, Texas 79430

Recent studies have suggested that the colonic H⁺,K⁺-ATPase (HK₂) can secrete either Na⁺ or H⁺ in exchange for K⁺. If correct, this view would indicate that the transporter could function as either a Na⁺ or a H⁺ pump. To investigate this possibility a series of experiments was performed using apical membranes from rat colon which were enriched in colonic H⁺,K⁺-ATPase protein. An antibody specific for HK₂ was employed to determine whether HK₂ functions under physiological conditions as a Na⁺-dependent or Na⁺-independent K⁺-ATPase in this same membrane fraction. K⁺-ATPase activity was measured as [-³²P]ATP hydrolysis. The Na⁺-dependent K⁺-ATPase accounted for approximately 80% of overall K⁺-ATPase activity and was characterized by insensitivity to Sch-28080 but partial sensitivity to ouabain. The Na⁺-independent K⁺-ATPase activity was insensitive to both Sch-28080 and ouabain. Both types of K⁺-ATPase activity substituted NH₄⁺ for K⁺ in a similar manner. Furthermore, our results demonstrate that when incubated with native distal colon membranes, the blocking antibody inhibited dramatically Na⁺-dependent K⁺-ATPase activity. Therefore, these data demonstrate that HK₂ can function in native distal colon apical membranes as a Na⁺-dependent K⁺-ATPase. Elucidation of the role of the pump as a transporter of Na⁺ versus H⁺ or NH₄⁺ versus K⁺ in vivo will require additional studies.

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