Purification and comparative studies of several mitochondrial aspartate aminotransferases from avian liver.
A new purification method has been developed which only exploits the chromatographic behaviour of avian liver mitochondrial aspartate aminotransferase enzymes (m-AAT), and permits a rapid isolation of the protein (4 days) in large quantities with high yield and low cost. m-AAT from turkey, chicken and quail livers have been isolated by chromatography on CM-Sepharose, Sephadex G-100 and 5' AMP-Sepharose using TEA-acetate buffer (pH 7.4), and specific activities (A.E.) of 311.6, 318.9, 320.1 I.U./mg respectively were obtained. Preparations were homogeneous as judged by various electrophoretic techniques and by size exclusion HPLC. The amino acid composition, Stokes Radius, subunit molecular weight and pI values have been determined and compared, finding no appreciable differences among them. In contrast, the absorption spectrum of the turkey enzyme differed from those of chicken and quail at both pH 7.4 and pH 5.0.
Department of Biochemistry, Faculty of Chemistry, University of Barcelona, Spain.
M
M
Cascante
A
A
Cortés
J
J
Bozal
D000818:Animals; D001219:Aspartate Aminotransferases / Q000302:isolation & purification*; D001717:Birds / Q000378:metabolism*; D002645:Chickens / Q000378:metabolism; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D008930:Mitochondria, Liver / Q000201:enzymology*; D008970:Molecular Weight; D011784:Quail / Q000378:metabolism; D013053:Spectrophotometry; D014422:Turkeys / Q000378:metabolism
D001219:Aspartate Aminotransferases