Kir6.2ΔC26 Channel Traffics to Plasma Membrane by Constitutive Exocytosis

Adenosine triphosphate （ATP）-sensitive K^＊ （KATP） channels regulate many cellular functions by coupling the metabolic state of the cell to the changes in membrane potential. Truncation of C-terminal 26 amino acid residues of Kir6.2 protein （Kir6.2ΔC26） deletes its endoplasmic reticulum retention signal, allowing functional expression of Kit6.2 in the absence of sulfonylurea receptor subunit, pEGFP-Kir6.2ΔC26 and pKir6.2ΔC26-IRES2-EGFP expression plasmids were constructed and transfected into HEK293 cells. We identified that Kir6.2ΔC26 was localized on the plasma membrane and trafficked to the plasmalemma by means of constitutive exocytosis of Kir6.2ΔC26 transport vesicles, using epi-fluorescence and total intemal reflection fluorescence microscopy. Our electrophysiological data showed that Kir6.2ΔC26 alone expressed KATP currents, whereas EGFP-Kir6.2ΔC26 fusion protein displayed no KATP channel activity.

Kir6.2DeltaC26 channel traffics to plasma membrane by constitutive exocytosis

Adenosine triphosphate (ATP)-sensitive K+ (KATP) channels regulate many cellular functions by coupling the metabolic state of the cell to the changes in membrane potential. Truncation of C-terminal 26 amino acid residues of Kir6.2 protein (Kir6.2DeltaC26) deletes its endoplasmic reticulum retention signal, allowing functional expression of Kir6.2 in the absence of sulfonylurea receptor subunit. pEGFP-Kir6.2DeltaC26 and pKir6.2DeltaC26-IRES2-EGFP expression plasmids were constructed and transfected into HEK293 cells. We identified that Kir6.2DeltaC26 was localized on the plasma membrane and trafficked to the plasmalemma by means of constitutive exocytosis of Kir6.2DeltaC26 transport vesicles, using epi-fluorescence and total internal reflection fluorescence microscopy. Our electrophysiological data showed that Kir6.2DeltaC26 alone expressed KATP currents, whereas EGFP-Kir6.2DeltaC26 fusion protein displayed no KATP channel activity.