Multiple sequences in the C terminus of MaxiK channels are involved in expression, movement to the cell surface, and apical localization

Apical expression of the large-conductance, calcium- and voltage-activated potassium (MaxiK) channel in the cortical collecting duct is responsible for flow-stimulated potassium secretion. Here, we identify two cytoplasmic regions controlling apical expression of the MaxiK channel. Disruption of the proximal region results in the intracellular retention of the MaxiK channel without affecting channel assembly, thereby reducing surface expression. Coexpression of the WT channel with this mutant results in a reduction of WT MaxiK channel at the cell surface. Our data indicate that this proximal region is necessary for export of the MaxiK channel from the endoplasmic reticulum as a way to assess the final assembly of the channel. Deletion of a more distal region disrupts apical sorting, resulting in a nonpolarized distribution of the channel without impairing its surface delivery. In summary, we have found that sequences of amino acids in the C terminus of the MaxiK channel operate after the channel is assembled into a multimer and play a role in its expression, movement to the cell surface, and apical localization

Basolateral membrane expression of a K(+) channel, Kir 2.3, is directed by a cytoplasmic COOH-terminal domain

The inwardly rectifying potassium channel Kir 2.3 is specifically targeted and expressed on the basolateral membrane of certain renal epithelial cells. In the present study, the structural basis for polarized targeting was elucidated. Deletion of a unique COOH-terminal domain produced channels that were mistargeted to the apical membrane, consistent with the removal of a basolateral membrane-sorting signal. By characterizing a series of progressively smaller truncation mutants, an essential targeting signal was defined (residues 431–442) within a domain that juxtaposes or overlaps with a type I PDZ binding motif (442). Fusion of the COOH-terminal structure onto CD4 was sufficient to change a random membrane-trafficking and expression pattern into a basolateral membrane one. Using metabolic labeling and pulse–chase and surface immunoprecipitation, we found that CD4-Kir2.3 COOH-terminal chimeras were rapidly and directly targeted to the basolateral membrane, consistent with a sorting signal that is processed in the biosynthetic pathway. Collectively, the data indicate that the basolateral sorting determinant in Kir 2.3 is composed of a unique arrangement of trafficking motifs, containing tandem, conceivably overlapping, biosynthetic targeting and PDZ-based signals. The previously unrecognized domain corresponds to a highly degenerate structure within the Kir channel family, raising the possibility that the extreme COOH terminus of Kir channels may differentially coordinate membrane targeting of different channel isoforms