Ho C Y, Barr L G, Clifford A J
Biochem Genet. 1979 Apr;17(3-4):209-21. doi: 10.1007/BF00498962.
The degree of relatedness among mammalian xanthine oxidases (XO) was determined by microcomplement fixation. Rabbit anti bovine milk XO serum was tested against xanthine oxidase (homologous protein) and against the heterologous proteins of bovine liver, monkey liver, rat liver, lactating cow serum, nonlactating cow serum, and steer serum. The indices of dissimilarity for the heterologous proteins were expressed as units of immunological distance and the percent sequence differences among these proteins inferred from the y = 5x relationship where y is immunological distance and x is percent sequence difference. Rat liver XO differed by approximately 27% in amino acid sequence from bovine milk XO. In order of increasing immunological distance from bovine milk XO, the sources of XO ranked as follows: lactating cow serum less than nonlactating cow serum less than steer serum = beef liver less than monkey liver less rat liver. The monkey ranked much closer than the rat in order of phylogenetic kinship to the cow. Starch gel electrophoresis of liver, milk, and serum showed that the milk and the serum contained only cationic forms of xanthine oxidase while all the liver samples tested contained cationic as well as anionic forms of the enzyme. The electrophoretic mobility properties of xanthine oxidase confirmed the polymorphic nature of the enzyme as revealed by the immunological data.
通过微量补体结合法测定了哺乳动物黄嘌呤氧化酶(XO)之间的亲缘程度。用兔抗牛乳XO血清检测黄嘌呤氧化酶(同源蛋白)以及牛肝、猴肝、大鼠肝、泌乳母牛血清、非泌乳母牛血清和公牛血清中的异源蛋白。异源蛋白的差异指数以免疫距离单位表示,并根据y = 5x关系推断这些蛋白之间的序列差异百分比,其中y为免疫距离,x为序列差异百分比。大鼠肝XO与牛乳XO的氨基酸序列差异约为27%。按照与牛乳XO免疫距离增加的顺序,XO的来源排序如下:泌乳母牛血清<非泌乳母牛血清<公牛血清 = 牛肉肝<猴肝<大鼠肝。在系统发育亲缘关系上,猴比大鼠更接近牛。对肝、乳和血清进行淀粉凝胶电泳显示,乳和血清中仅含有阳离子形式的黄嘌呤氧化酶,而所有检测的肝样本中均含有阳离子和阴离子形式的该酶。黄嘌呤氧化酶的电泳迁移特性证实了该酶如免疫数据所示的多态性本质。