PURIFICATION AND PROPERTIES OF THE PLASMA-MEMBRANE H+-TRANSLOCATING ADENOSINE-TRIPHOSPHATASE OF PHASEOLUS-MUNGO L ROOTS
PURIFICATION AND PROPERTIES OF THE PLASMA-MEMBRANE H+-TRANSLOCATING ADENOSINE-TRIPHOSPHATASE OF PHASEOLUS-MUNGO L ROOTS
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DOI：
10.1104/pp.80.4.818
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发表时间：
1986-04-01
期刊：
影响因子：
7.4
通讯作者：
KASAMO, K
中科院分区：
文献类型：
--
作者：
KASAMO, K
绿豆（Phaseolus mungo L.）根的质膜ATP通过阴离子洗涤剂脱氧胆酸盐（DOC）和两性离子洗涤剂（zwitter剂3-14）两步溶出：(a)松散结合的膜蛋白通过0.1% DOC处理去除；(b)在1% DOC存在下，用0.1%的两性溶剂溶解atp酶；(c)通过甘油梯度（45-70%）离心进一步纯化溶解的物质。通常，大约10%的atp酶活性恢复，比活性增加约11倍。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳表明，甘油梯度峰段含有Mr = 105,000, 67,000和57,000道尔顿的三种主要多肽。纯化后的atp酶在pH最佳、底物特异性、抑制剂敏感性和离子刺激方面与膜结合的atp酶基本相同。
The plasma membrane ATP of mung bean (Phaseolus mungo L.) roots has been solubilized with a two-step procedure using the anionic detergent, deoxycholate (DOC) and the zwitterionic detergent, zwittergent 3-14 as follows: (a) loosely bound membrane proteins are removed by treatment with 0.1% DOC; (b) The ATPase is solubilized with 0.1% zwittergent in the presence of 1% DOC; (c) the solubilized material is further purified by centrifugation through a glycerol gradient (45-70%). Typically, about 10% of the ATPase activity is recovered, and the specific activity increases about 11-fold. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that the peak fraction from the glycerol gradient contains three major polypeptides of Mr = 105,000, 67,000, and 57,000 daltons. The properties of the purified ATPase are essentially the same as those of membrane-bound ATPase, with respect to pH optimum, substrate specificity, inhibitor sensitivity, and ion stimulation.