Polarized membrane distribution of potassium-dependent ion pumps in epithelial cells: Different roles of the N-glycans of their β subunits

Abstract

The Na,K-ATPases and the H,K-ATPases are two potassium-dependent homologous heterodimeric P2-type pumps that catalyze active transport of Na+ in exchange for K+ (Na,K-ATPase) or H+ in exchange for K+ (H,K-ATPase). The ubiquitous Na,K-ATPase maintains intracellular ion balance and membrane potential. The gastric H,K-ATPase is responsible for acid secretion by the parietal cell of the stomach. Both pumps consist of a catalytic alpha-subunit and a glycosylated beta-subunit that is obligatory for normal pump maturation and trafficking. Individual N-glycans linked to the beta-subunits of the Na,K-ATPase and H,K-ATPase are important for stable membrane integration of their respective alpha subunits, folding, stability, subunit assembly, and enzymatic activity of the pumps. They are also essential for the quality control of unassembled beta-subunits that results in either the exit of the subunits from the ER or their ER retention and subsequent degradation. Overall, the importance of N-glycans for the maturation and quality control of the H,K-ATPase is greater than that of the Na,K-ATPase. The roles of individual N-glycans of the beta-subunits in the post-ER trafficking, membrane targeting and plasma membrane retention of the Na,K-ATPase and H,K-ATPase are different. The Na,K-ATPase beta1-subunit is the major beta-subunit isoform in cells with lateral location of the pump. All three N-glycans of the Na,K-ATPase beta1-subunit are important for the lateral membrane retention of the pump due to glycan-mediated interaction between the beta1-subunits of the two neighboring cells in the cell monolayer and cytosolic linkage of the alpha-subunit to the cytoskeleton. This intercellular beta1-beta1 interaction is also important for formation of cell-cell contacts. In contrast, the N-glycans unique to the Na,K-ATPase beta2-subunit,which has up to eight N-glycosylation sites, contain apical sorting information. This is consistent with the apical location of the Na,K-ATPase in normal and malignant epithelial cells with high abundance of the beta2-subunit. Similarly, all seven N-glycans of the gastric H,K-ATPase beta-subunit determine apical sorting of this subunit.