title: Electrophoretic and kinetic characterization of three variants of soluble cytoplasmic L-alanine:2-oxoglutarate aminotransferase in human liver tissue.
abstract: Three common variants of soluble cytoplasmic L-alanine:2-oxoglutarate aminotransferase (ALT, EC 2.6.1.2), sALT 1, 2-1 and 2, were isolated from normal human liver, and characterized by electrophoretic and kinetic analyses. The isoelectric point of sALT 1 was pH 6.45. sALT 2-1 was focused into three bands with pl 6.1, 6.2 and 6.45; sALT was focused into one band with pl 6.1. The electrophoretic mobilities of sALTs altered to the fast beta-globulin fraction after aging or papain treatment. Ammonia was produced during the latter, and the altered migration was considered to be caused by deamidation of sALT. The relative molecular mass of each of the enzymes was 110,000. Minor differences in the apparent Km values among the multiple forms for both L-alanine and 2-oxoglutarate were observed after incubation with 100 mumol/L of pyridoxal phosphate (PALP). PALP stimulation of the enzyme activities was also different. sALT 1 was more stable than sALT 2-1 and 2 after heat and urea treatments. In human sera from 1065 adult Japanese, sALT 2-1, a heterozygote form of sALT 1 and 2, was dominant.
authors.affiliation: Division of Clinical Laboratories, Kurashiki Central Hospital, Okayama, Japan.
authors.forename: F
authors.initials: F
authors.lastname: Kanemitsu
authors.forename: I
authors.initials: I
authors.lastname: Kawanishi
authors.forename: J
authors.initials: J
authors.lastname: Mizushima
authors.forename: T
authors.initials: T
authors.lastname: Okigaki
mesh_terms: D000410:Alanine Transaminase / Q000302:isolation & purification / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology; D004586:Electrophoresis; D004789:Enzyme Activation; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D011732:Pyridoxal Phosphate / Q000494:pharmacology
chemical_list: D007527:Isoenzymes; D011732:Pyridoxal Phosphate; D000410:Alanine Transaminase