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腺苷脱氨酶是机体重要腺苷代谢酶之一,能催化腺苷嘌呤环上6-位氨基脱氨,从而生成肌苷的氨解反应,与腺苷激酶、5′-核苷酸酶及 S-腺苷同型半胱氨酸水解酶共同使机体腺苷水平保持相对稳定。文献报道人体组织中的腺苷脱氨酶分子有多种形式,其表观分子量分别为36000、114000、200000和298000。本文报告一种用亲和层析法,从人红细胞胞浆液中分离腺苷脱氨酶的新的简便快速方法。所采用的亲和层析胶,为 N~6-(6-氨己基)腺苷-Sepharose4 B 凝胶,是由N~6-(6-氨己基)腺苷与经溴化氰活化的 Sepharose4 B 胶偶联而成。人红细胞胞浆液中的腺苷脱氨酶,经两次亲和层析分离后,比活性提高了197倍,酶活性回收率为49%。高效液相凝胶排阻层析分析的结果表明,分离出的酶蛋白制剂含有表观分子量分别为36000、72000、114000和64000的四种蛋白质组分。其中前三种组分的表观分子量与文献报道基本相符。
Adenosine deaminase is one of the important adenosine metabolic enzymes in the body and can catalyze the deamination of the amino group at the 6-position of adenosine ring to form the aminolysis reaction of inosine and adenosine kinase, 5’-nucleotidase And S-adenosyl cysteine hydrolase together to maintain the level of adenosine in the body relatively stable. Reported in the literature of human tissue adenosine deaminase molecules in many forms, the apparent molecular weight were 36000,114,000,200,000 and 298,000. This paper reports a novel and rapid method for the isolation of adenosine deaminase from human erythrocyte cyst by affinity chromatography. The affinity chromatography gel used was N ~ 6- (6-aminohexyl) adenosine-Sepharose 4B gel and consisted of N ~ 6- (6-aminohexyl) adenosine and Sepharose 4 activated by cyanogen bromide B plastic coupling. The adenosine deaminase in human erythrocyte cytosolic fluid was 197 times more specific and the recovery of enzyme activity was 49% after two affinity chromatography. The results of high performance liquid chromatography-gel exclusion chromatography showed that the isolated enzyme protein preparation contained four protein components with apparent molecular weights of 36000, 72000, 114000 and 64000, respectively. The first three components of the apparent molecular weight and reported in the literature basically.