Purzyc L, Otrebski E, Goluda M, Kwiatkowska J
Neoplasma. 1984;31(3):307-14.
Alkaline phosphatase from human uterine myoma was purified to homogeneity by butanol extraction, acetone precipitation, column chromatography on DEAE-cellulose, QAE-Sephadex and Con-A-Sepharose, gel filtration on Sephadex G-200 and preparative electrophoresis. The relative molecular mass (145 000), subunit mass (35 000), neutral sugars (7 micrograms/mg) and sialic acid (1 microgram/mg) content were estimated. pH optimum of the enzyme activity was 10.0-10.4, and Km for p-nitrophenyl phosphate was 0.23 mM. The enzyme was inhibited by Zn2+, phosphate, fluoride, EDTA and by treatment with neuraminidase. Mg2+ activated alkaline phosphatase and showed protective effect towards inhibition by EDTA and Zn2+. The uterine muscle alkaline phosphatase was also purified. It showed lower specific activity than myoma phosphatase, higher molecular mass (160 000) and subunit mass (76 400), higher neutral sugar and sialic acid content.
通过丁醇萃取、丙酮沉淀、DEAE - 纤维素柱色谱、QAE - 葡聚糖凝胶柱色谱、伴刀豆球蛋白A - 葡聚糖凝胶柱色谱、Sephadex G - 200凝胶过滤和制备性电泳，将人子宫肌瘤碱性磷酸酶纯化至同质。测定了其相对分子质量（145000）、亚基质量（35000）、中性糖含量（7微克/毫克）和唾液酸含量（1微克/毫克）。该酶活性的最适pH为10.0 - 10.4，对磷酸对硝基苯酯的Km值为0.23毫摩尔。该酶受到锌离子、磷酸盐、氟化物、乙二胺四乙酸（EDTA）的抑制，并且经神经氨酸酶处理后也会受到抑制。镁离子激活碱性磷酸酶，并对EDTA和锌离子的抑制作用具有保护作用。子宫肌碱性磷酸酶也得到了纯化。它的比活性低于肌瘤磷酸酶，相对分子质量更高（160000），亚基质量更高（76400），中性糖和唾液酸含量更高。
