猪肾和人胎盘二胺氧化酶的纯化及某些性质

Purification and some properties of diamine oxidase from pig kidney and human placenta.

Hata A

Bull Tokyo Med Dent Univ. 1976 Jun;23(2):63-70.

Abstract

A homogeneous preparation of pig kidney diamine oxidase was obtained by polyacrylamide gel electrophoresis and immunodiffusion. Its molecular weight was estimated as 170,000 by Sephadex G-200 column chromatography and as 220,000 by SDS-polyacrylamide gelelectrophoresis. After mercaptoethanol cleavage of disulfide bonding, the molecular weight of the subunit was 130,000. 2. Little contamination was observed in the preparation of human placenta by SDS-polyacrylamide gel electrophoresis and immunoelectrophoresis and the molecular weight was calculated as 30008000 and 280,000 by the extrapolation from the calibration curve of Sephadex G-200 column chromatography and SDS-polyacrylamide gel electrophoresis, respectively. The subunit was 170,000. 3. By immunodiffusion, immunoelectrophoresis and inhibition test of the enzyme activity using a heterologous antiserum, a good cross-reactivity was observed between the anti-human diamine oxidase serum and pig diamine oxidase. 4. The activities of the two enzymes were not almost inhibited by homologous and heterologous antisera.

摘要

通过聚丙烯酰胺凝胶电泳和免疫扩散获得了猪肾二胺氧化酶的均一制剂。用葡聚糖凝胶G - 200柱色谱法估计其分子量为170,000，用SDS - 聚丙烯酰胺凝胶电泳法估计为220,000。在二硫键经巯基乙醇裂解后，亚基的分子量为130,000。2. 通过SDS - 聚丙烯酰胺凝胶电泳和免疫电泳观察到，人胎盘制剂中的污染很少，通过葡聚糖凝胶G - 200柱色谱校准曲线外推法计算分子量为30008000，通过SDS - 聚丙烯酰胺凝胶电泳法计算为280,000。亚基为170,000。3. 通过免疫扩散、免疫电泳以及使用异种抗血清对酶活性进行抑制试验，观察到抗人二胺氧化酶血清与猪二胺氧化酶之间具有良好的交叉反应性。4. 这两种酶的活性几乎未被同源和异种抗血清抑制。