Differential localization of human nongastric H(+)-K(+)-ATPase ATP1AL1 in polarized renal epithelial cells.
The human H(+)-K(+)-ATPase, ATP1AL1, belongs to the subgroup of nongastric, K(+)-transporting ATPases. In concert with the structurally related gastric H(+)-K(+)-ATPase, it plays a major role in K(+) reabsorption in various tissues, including colon and kidney. Physiological and immunocytochemical data suggest that the functional heteromeric ion pumps are usually found in the apical plasma membranes of renal epithelial cells. However, the low expression levels of characteristic nongastric ion pumps makes it difficult to verify their spatial distribution in vivo. To investigate the sorting behavior of ATP1AL1, we expressed this pump by stable transfection in MDCK and LLC-PK(1) renal epithelial cell lines. Stable interaction of ATP1AL1 with either the endogenous Na(+)-K(+)-ATPase beta-subunit or the gastric H(+)-K(+)-ATPase beta-subunit was tested by confocal immunofluorescence microscopy and surface biotinylation. In cells transfected with ATP1AL1 alone, the alpha-subunit accumulated intracellularly, consistent with its inability to assemble and travel to the plasma membrane with the endogenous Na(+)-K(+)-ATPase beta-subunit. Cotransfection of ATP1AL1 with the gastric H(+)-K(+)-ATPase beta-subunit resulted in plasma membrane localization of both pump subunits. In cotransfected MDCK cells the heteromeric ion pump was predominantly polarized to the apical plasma membrane. Functional expression of ATP1AL1 was confirmed by (86)Rb(+) uptake measurements. In contrast, cotransfected LLC-PK(1) cells accumulate ATP1AL1 at the lateral membrane. The distinct polarization of ATP1AL1 indicates that the alpha-subunit encodes sorting information that is differently interpreted by cell type-specific sorting mechanisms.
Reinhardt
J
J
Department of Physiology, University of Münster, 48149 Münster, Germany.
Grishin
A V
AV
Oberleithner
H
H
Caplan
M J
MJ
D000818:Animals; D016764:Cell Polarity / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology / Q000201:enzymology*; D005455:Fluorescent Antibody Technique; D015971:Gene Expression Regulation, Enzymologic; D017506:H(+)-K(+)-Exchanging ATPase / Q000032:analysis* / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000201:enzymology*; D018374:LLC-PK1 Cells; D018613:Microscopy, Confocal; D013270:Stomach / Q000201:enzymology; D013552:Swine; D014162:Transfection
D017506:H(+)-K(+)-Exchanging ATPase