通过亲和色谱法分离ADP-核糖基转移酶。
Isolation of ADP-ribosyltransferase by affinity chromatography.
作者信息
Burtscher H J, Auer B, Klocker H, Schweiger M, Hirsch-Kauffmann M
出版信息
Anal Biochem. 1986 Feb 1;152(2):285-90. doi: 10.1016/0003-2697(86)90410-0.
An affinity adsorbent for ADP-ribosyltransferase (EC 2.4.2.30) has been synthesized by coupling 3-aminobenzamide to Sepharose 4B. Using this material, ADP-ribosyltransferase from human placenta has been purified from crude extract to homogeneity within a few hours. The enzyme has an apparent Km for NAD+ of 52 microM. Its molecular mass is 115,000 as determined by gel electrophoresis. The enzyme is DNA dependent and stimulated by histone, its temperature optimum is at 25 degrees C, and its pH optimum is around pH 9. alpha-NAD+, thymidine, caffeine, theophylline, theobromine, 3-methoxybenzamide, and nicotinamide inhibit the enzyme. Purification of ADP-ribosyltransferases from horse, rat, and chicken liver was also achieved with the method described.
通过将3-氨基苯甲酰胺偶联到琼脂糖4B上，合成了一种用于ADP-核糖基转移酶（EC 2.4.2.30）的亲和吸附剂。使用这种材料，几小时内就将人胎盘来源的ADP-核糖基转移酶从粗提物中纯化至均一。该酶对NAD +的表观Km为52微摩尔。通过凝胶电泳测定，其分子量为115,000。该酶依赖于DNA并受组蛋白刺激，其最适温度为25℃，最适pH约为pH 9。α-NAD +、胸苷、咖啡因、茶碱、可可碱、3-甲氧基苯甲酰胺和烟酰胺可抑制该酶。用所述方法也实现了从马、大鼠和鸡肝脏中纯化ADP-核糖基转移酶。
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