皱纹盘鲍中肠腺两种酸性磷酸酶的纯化及性质

Purification and properties of two acid phosphatases from midgut glands of abalone Haliotis discus.

作者信息

Yoshida H, Sagami H, Oikawa S

出版信息

J Biochem. 1977 May;81(5):1447-54.

Abstract

Midgut glands of abalone Haliotis discus contained two acid phosphatases [orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2] separable by phosphocellulose column chromatography. They were designated as acid phosphatases I and II in order of elution and were purified 99- and 290-fold, respectively. Purified acid phosphatase II was nearly homogeneous as judged by polyacrylamide gel electrophoresis. The substrate specificity of acid phosphatase I was narrow, whereas that of acid phosphatase II was broad. Good substrates for acid phosphatase I included p-nitrophenyl phosphate, phosphoenolpyruvate, inorganic pyrophosphate, and nucleoside di- and triphosphates. The acid phosphatases did not require any metal ion for maximum activity and were inhibited by Zn2+, Cu2+ and Hg2+. Fluoride and arsenate were potent inhibitors of both enzymes. The pH optima of acid phosphatases I and II were 5.9 and 5.5, respectively. The molecular weights of acid phosphatases I and II were estimated to be 28,000 and 100,000, respectively, by gel filtration on Sephadex G-100. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis suggested that acid phosphatase II consists of two identical subunits.

摘要

皱纹盘鲍的中肠腺含有两种酸性磷酸酶[正磷酸单酯磷酸水解酶（最适酸性），EC 3.1.3.2]，可通过磷酸纤维素柱色谱分离。按照洗脱顺序，它们分别被命名为酸性磷酸酶I和II，纯化倍数分别为99倍和290倍。通过聚丙烯酰胺凝胶电泳判断，纯化后的酸性磷酸酶II几乎是均一的。酸性磷酸酶I的底物特异性较窄，而酸性磷酸酶II的底物特异性较宽。酸性磷酸酶I的良好底物包括对硝基苯磷酸酯、磷酸烯醇丙酮酸、无机焦磷酸以及核苷二磷酸和三磷酸。这些酸性磷酸酶在最大活性时不需要任何金属离子，并且受到Zn2+、Cu2+和Hg2+的抑制。氟化物和砷酸盐是这两种酶的有效抑制剂。酸性磷酸酶I和II的最适pH分别为5.9和5.5。通过在Sephadex G - 100上进行凝胶过滤，估计酸性磷酸酶I和II的分子量分别为28,000和100,000。十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳表明酸性磷酸酶II由两个相同的亚基组成。