title: Purification and characterization of porcine skeletal muscle aminopeptidase T, a novel metallopeptidase homologous to leukotriene A4 hydrolase.
abstract: A novel aminopeptidase, Aminopeptidase T (APase T), was purified from porcine skeletal muscle following successive column chromatography: twice on DEAE-cellulose, hydroxyapatite, and Sephacryl S-200 HR using Leu-β-naphthylamide (LeuNap) as a substrate. The molecular mass of the enzyme was 69 kDa on SDS-PAGE. The optimum pH towards LeuNap of the enzyme was about 7. The enzyme activity was strongly inhibited by bestatin and was negatively affected by ethylenediaminetetraacetic acid (EDTA). Chlorine-activated APase T liberated Leu, Ala, Met, Pro, and Arg from Nap derivatives. The APase T gene consisted of an ORF of 1,836 bp encoding a protein of 611 amino acid residues. The APase T was highly homologous to bovine, human, and mouse Leukotriene A(4) hydrolase (LTA(4)H), a bifunctional enzyme which exhibits APase and epoxide hydrolase activity.
authors.affiliation: Graduate School of Biosphere Science, Hiroshima University, Hiroshima, Japan.
authors.forename: Mohammed Alamgir
authors.initials: MA
authors.lastname: Sarker
authors.forename: Shinji
authors.initials: S
authors.lastname: Matsuda
authors.forename: Osamu
authors.initials: O
authors.lastname: Mizutani
authors.forename: Shengbin
authors.initials: S
authors.lastname: Rao
authors.forename: Koshiro
authors.initials: K
authors.lastname: Migita
authors.forename: Nami
authors.initials: N
authors.lastname: Goto-Yamamoto
authors.forename: Haruyuki
authors.initials: H
authors.lastname: Iefuji
authors.forename: Toshihide
authors.initials: T
authors.lastname: Nishimura
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002713:Chlorine / Q000494:pharmacology; D002845:Chromatography; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004851:Epoxide Hydrolases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D007930:Leucine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018482:Muscle, Skeletal / Q000737:chemistry / Q000201:enzymology*; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D034421:Sus scrofa / Q000378:metabolism
chemical_list: D000596:Amino Acids; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D002713:Chlorine; C040135:leucine-beta-naphthylamide; D004492:Edetic Acid; D004851:Epoxide Hydrolases; D000626:Aminopeptidases; D007930:Leucine; C012211:ubenimex; C043134:leukotriene A4 hydrolase