title: An enkephalin-degrading aminopeptidase from rat brain catalyzes the hydrolysis of a neuropeptide, kyotorphin (L-Tyr-L-Arg).
abstract: We studied the hydrolysis of a neuropeptide kyotorphin (L-Tyr-L-Arg) by an enkephalin-degrading aminopeptidase purified from cytosol of rat brain in vitro. The purified enzyme was homogeneous as judged by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE), gel filtration and isoelectric focusing. The aminopeptidase with an apparent molecular weight (Mr) = 98000 catalyzed the hydrolysis of Leu- and Met-enkephalins with Km values of 125 and 142 microM, respectively. The enzyme activity was inhibited by bestatin, amastatin and puromycin but not by pepstatin, leupeptin and phenylmethanesulfonyl fluoride (PMSF). Kyotorphin was degraded by the aminopeptidase at pH 7.0, and the Vmax and Km values were 9.2 mumol/min/mg protein and 95 microM, respectively. The Km value for kyotorphin was compatible to those for Leu- and Met-enkephalins. Taken together, these results suggest a possible involvement of the enkephalin-degrading aminopeptidase in cytosolic degradation of kyotorphin in neuronal cells of rat brain.
authors.affiliation: Faculty of Pharmacy & Pharmaceutical Sciences, Fukuyama University, Hiroshima, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Akasaki
authors.forename: H
authors.initials: H
authors.lastname: Tsuji
mesh_terms: D000626:Aminopeptidases / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000201:enzymology*; D004723:Endorphins / Q000378:metabolism*; D004745:Enkephalins / Q000378:metabolism*; D006868:Hydrolysis; D051381:Rats
chemical_list: D004723:Endorphins; D004745:Enkephalins; C019448:kyotorphin; D000626:Aminopeptidases