title: Purification and characterization of acid phosphatase in rat liver lysosomal contents.
abstract: Acid phosphatase in rat liver lysosomal contents, C-APase I, was purified about 5,700-fold over the homogenate with 8.0% recovery, to apparent homogeneity as determined from the pattern on polyacrylamide gel electrophoresis in the presence and in the absence of SDS. The purification procedures included; preparation of crude lysosomal contents, DEAE-Sephacel ion exchange chromatography, hydroxylapatite chromatography, and gel filtration with Sephacryl S-300. The enzyme is composed of three identical subunits with an apparent molecular weight of 48K. The enzyme contains about 11% carbohydrate and the carbohydrate moiety was composed of mannose, fucose, N-acetylglucosamine, and N-acetylgalactosamine in a molar ratio of 20:3:11:1. Sialic acid was not detected in the enzyme. Antisera against the purified C-APase I were raised in goat and the C-APase I was rapidly purified with high yield (10%) by using the specific antibodies coupled to Sepharose 6B.
authors.affiliation: Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka.
authors.forename: M
authors.initials: M
authors.lastname: Himeno
authors.forename: H
authors.initials: H
authors.lastname: Koutoku
authors.forename: H
authors.initials: H
authors.lastname: Tsuji
authors.forename: K
authors.initials: K
authors.lastname: Kato
mesh_terms: D000135:Acid Phosphatase / Q000276:immunology / Q000302:isolation & purification*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000917:Antibody Formation; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006023:Glycoproteins; D008099:Liver / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology*; D046911:Macromolecular Substances; D008297:Male; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D000596:Amino Acids; D006023:Glycoproteins; D046911:Macromolecular Substances; D000135:Acid Phosphatase