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Volume 271, Number 47, Issue of November 22, 1996 pp. 29759-29763
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Functional Expression of the Colonic H+,K+-ATPase alpha -Subunit
PHARMACOLOGIC PROPERTIES AND ASSEMBLY WITH X+,K+-ATPase beta -SUBUNITS

(Received for publication, August 6, 1996)

Juan Codina , Bruce C. Kone , Juan T. Delmas-Mata and Thomas D. DuBose Jr.

From the Departments of Internal Medicine and Integrative Biology, The University of Texas Medical School at Houston, Houston, Texas 77030

The functional and pharmacological properties of the alpha -subunit of the colonic H+,K+-ATPase (alpha C) were studied in Xenopus laevis oocytes. alpha C was injected with different rat beta -subunits, the beta -subunit of the gastric H+,K+-ATPase (beta G, the only H+,K+-ATPase beta -subunit identified in rat), or the beta 1-subunit of the Na+,K+-ATPase (beta 1) (associated with the basolateral Na+,K+-ATPase, but also expressed in the epithelial apical membranes of rat distal colon) (Marxer, A., Stieger, B., Quarini, A., Kashgarian, M., and Hauri, H. P. (1989) J. Cell Biol. 109, 1057-1069). The effect of the different beta -subunits was studied by measuring 86Rb+ uptake (a K+ congener) in the presence or absence of Sch-28080 and ouabain. Significant Na+-independent 86Rb+ uptake was observed only when alpha C was coexpressed with one of the beta -subunits. The expressed alpha Cbeta 1 and alpha Cbeta G complexes were not inhibited by Sch-28080, were only partially sensitive to ouabain (IC50 = 400-600 µM, in the presence of external 1 mM KCl), and exhibited comparable K+ activation kinetics. Coexpression of alpha C with epitope-tagged beta G or beta 1, followed by immunopurification of the alpha beta complexes, confirmed stable assembly of alpha Cbeta G and alpha Cbeta 1 complexes. Since the beta 1-subunit, but not the alpha 1-subunit, of Na+,K+-ATPase is expressed in the apical membrane of rat colonocytes, our data support the view that, in rat distal colon, the beta 1-subunit may play a surrogate role as the beta -subunit for the colonic H+,K+-ATPase.


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