E2P state stabilization by the N-terminal tail of the H,K-ATPase beta-subunit is critical for efficient proton pumping under in vivo conditions.

ABSTRACT: The catalytic alpha-subunits of Na,K- and H,K-ATPase require an accessory beta-subunit for proper folding, maturation, and plasma membrane delivery but also for cation transport. To investigate the functional significance of the beta-N terminus of the gastric H,K-ATPase in vivo, several N-terminally truncated beta-variants were expressed in Xenopus oocytes, together with the S806C alpha-subunit variant. Upon labeling with the reporter fluorophore tetramethylrho da mine-6-maleimide, this construct can be used to determine the voltage-dependent distribution between E(1)P/E(2)P states. Whereas the E(1)P/E(2)P conformational equilibrium was unaffected for the shorter N-terminal deletions betaDelta4 and betaDelta8, we observed significant shifts toward E(1)P for the two larger deletions betaDel

SUBMITTER: Durr KL 

PROVIDER: S-EPMC2740441 | biostudies-literature | 2009 Jul

REPOSITORIES: biostudies-literature