Boyle F A, Cook N D, Peters T J
Division of Clinical Cell Biology, MRC Clinical Research Centre, Harrow, Middlesex, UK.
Clin Chim Acta. 1988 Feb 15;171(2-3):187-96. doi: 10.1016/0009-8981(88)90144-1.
Galactosyltransferase was isolated from human malignant ascites by ammonium sulphate precipitation, alpha-lactalbumin affinity chromatography and by the removal of contaminating immunoglobulins with protein A chromatography. Two isoforms of galactosyltransferase were separated by DEAE chromatography, and partially characterised by gel filtration, non-denaturing polyacrylamide gel electrophoresis, concanavalin A binding and enzyme kinetics. The two forms isolated were compared with the isoenzymes previously described and the biochemical basis for the difference between the two forms discussed.
通过硫酸铵沉淀、α-乳白蛋白亲和层析以及用蛋白A层析去除污染的免疫球蛋白,从人恶性腹水中分离出半乳糖基转移酶。通过DEAE层析分离出半乳糖基转移酶的两种同工型,并通过凝胶过滤、非变性聚丙烯酰胺凝胶电泳、伴刀豆球蛋白A结合和酶动力学对其进行部分表征。将分离出的两种形式与先前描述的同工酶进行比较,并讨论了两种形式之间差异的生化基础。