Analysis of the interaction between human kidney anion exchanger 1 and kanadaptin using yeast two-hybrid systems

Abstract

Abstract Kidneyanionexchangeradaptorprotein(Kanadaptin)isaproteinwhichinteractswiththecytoplasmicN-terminaldo-main of kidney anion exchanger 1 (kAE1) and was first detected in mice using the yeast two-hybrid system and wasalso found to co-localize with kAE1 in rabbit -intercalated cells. Impaired trafficking of human kAE1 can result in thekidney disease-distal renal tubular acidosis (dRTA), and defective interaction between human kAE1 and kanadaptinmay cause this trafficking impairment and be the basis for dRTA pathogenesis. However, it is unknown whetherkAE1canreallyinteractwithkanadaptininhumans.WehavethusinvestigatedtheinteractionbetweenhumankAE1and human kanadaptin by using both Gal4 and LexA yeast two-hybrid systems. It was found that co-expression ofGal4DBD fused to the cytoplasmic N-terminal domain of kAE1 and Gal4AD fused to kanadaptin could not activatethe transcription of the ADE2 , HIS3 and lacZ reporters in the Gal4 system. A similar result was obtained for the inter-action between B42AD fused to the cytoplasmic N-terminal domain of kAE1 and LexA fused to kanadaptin in activa-tion of