THE PHOSPHINOUS ACID (CF3)2POH AND THE DIPHOSPHOXANE (CF3)2POP(CF3)21
THE PHOSPHINOUS ACID (CF3)2POH AND THE DIPHOSPHOXANE (CF3)2POP(CF3)21
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亚磷酸 (CF3)2POH 和二磷氧烷 (CF3)2POP(CF3)21
DOI：
10.1021/ja01491a062
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发表时间：
1960
影响因子：
15
通讯作者：
A. Burg
中科院分区：
文献类型：
--
作者：
J. E. Griffiths;A. Burg
猪肾的碱性磷酸酶存在于微粒体 2 的核糖核蛋白颗粒中，并通过蛋白水解作为活性片段释放，2·3 已通过应用先前描述的程序 3 加上 Ecteola 纤维素离子交换色谱法，在明显均质的条件下进行分离。 4 在典型的纯化中，活性为 2,000,000 单位2，比活性超过每毫克 100,000 单位。将总氮（微量凯氏定氮）置于 Ec-teola 5 X 100 cm 柱上。用水彻底洗涤后，用pH 9、0.01至0.05M梯度的乙酸钡在10升溶液中洗脱。活性材料对称地洗脱到接近 0.03 M，并通过钡程序 3 进行浓缩，得到约 1,500,000 单位的材料，基于总 N，其比活性为 295,000 至 310,000。在 Ecteola-纤维素、Deae-纤维素4 或 Dowex-23 上进行再色谱，纸电泳和纸色谱（乙醇-1 M 铵）醋酸盐，70-30）显示 280 µ 处的吸光度没有与活性分离。当未稀释测试时，该材料不含肽酶和二酯酶活性，氨基酸对活性没有影响，并且在任何时候都不可能证明除镁离子之外的可透析辅因子。紫外吸光度是蛋白质的特征，最大为278，最小为250μ。然而，在用稀酸（100℃下0.1至1.0M）处理的过程中，发现吸光度显着增加，并且在2小时结束时，278℃下的吸光度几乎增加了三倍。存在还原材料6、茚三酮反应材料6 以及与磷钼酸盐反应的材料的平行释放。 7 纸色谱法（丙醇-水，80-20）将磷钼酸盐和茚三酮反应物质与茚三酮阴性但邻苯二甲酸氢苯胺阳性8（棕色）物质分离，其 Ri 与葡萄糖相同。 Dische9 半胱氨酸方法中未经处理的材料给出
Alkaline phosphatase of swine kidney, found in the ribonucleoprotein particles of the microsomes2 and released as an active fragment by proteolysis, 2· 3 has been isolated in an apparently homogeneous condition by application of the procedures pre-viously described3 plus ion-exchange chromatography with Ecteola cellulose. 4 In a typical puri-fication 2,000,000 units activity2 with a specific activity of over 100,000 units per mg. total N (micro-Kjeldahl) was placed on a column of Ec-teola 5 X 100 cm. and, after thorough washing with water, was eluted with a gradient of barium acetate at pH 9 varying from 0.01 to 0.05 M in 10 liters of solution. The active material was eluted sym-metrically near 0.03 M and was concentrated by the barium procedure3to yield about 1,500,000 units material with a specific activity of 295,000 to 310,000 on the basis of total N. Rechromatography on Ecteola-cellulose, on Deae-cellulose4 or on Dowex-23, paper electrophoresis and paper chromatography (ethanol-1 M ammonium ace-tate, 70-30) revealed no dissociation of absorbancy at 280 µ from activity. The material was free of peptidase and diesterase activity when tested un-diluted, amino acids were without effect on the activity and at no time was it possible to demonstrate dialyzable cofactorsother than magnesium ion. The absorbancy in the ultraviolet was charac-teristic of protein with a maximum at 278 and a minimum at 250 mu. However, in the course of treatment with dilute acid (0.1 to 1.0 M at 100) the absorbancy was found to increase remarkably and, at the end of 2 hr., the absorbancyat 278 was nearly tripled. There was a parallel release of reducing material, 6 of ninhydrin reactive material, 6 and of material reacting with phosphomolybdate. 7 Paper chromatography (propanol-water, 80-20) separated a phosphomolybdate and ninhydrin reac-tive material from a ninhydrin negative but aniline hydrogen phthalate positive8 (brown color) mate-rial with the same Ri as glucose. The untreated material in the cysteine methods of Dische9 gave