ATP‐dependent transport of amphiphilic cations across the hepatocyte canalicular membrane mediated by mdr1 P‐glycoprotein

Abstract
The ATP‐dependent transport of the three 3H‐labeled, amphiphilic cations quinidine, N‐(n‐pentyl)‐quinidinium, and N‐(4',4'‐azo‐n‐pentyl) ‐21‐deoxyajmalinium was studied in rat canalicular plasma membrane vesicles. N‐Alkylation of quinidine with an n‐pentyl residue resulted in a permanently charged cationic substrate for ATP‐dependent transport which exhibited a 10‐fold higher transport rate relative to quinidine. The K m value was 0.4 μM for N‐(n‐pentyl)‐quinidinium and 5 μM for quinidine. The permanently cationic and photolabile derivative of ajmaline, N‐(4',4'‐azo‐n‐pentyl)‐21‐deoxyajmalinium, was also an efficient substrate and served to label canalicular membrane proteins with molecular masses of 143 kDa and 108 kDa. ATP‐dependent transport of the permanently charged amphiphilic cations was inhibited by the P‐glycoprotein inhibitors and substrates quinidine, verapamil, and daunorubicin. The data demonstrate that N‐alkylation of quinidine and ajmaline results in most efficient substrates for mdr1 P‐glycoprotein‐mediated ATP‐dependent transport.