植物28kD蛋白的纯化及部分性质研究

经丙酮粉、硫酸铵分级沉淀和阴离子交换层析等方法，首次从玉米花粉细胞质粗提物中纯化了一种具有ＡＴＰａｓｅ活性的可溶性蛋白。ＳＤＳ－ＰＡＧＥ测得分子量为２８ｋＤ，ＩＥＦ－ＰＡＧＥ测得等电点为８．３。免疫印迹鉴定结果表明该酶蛋白与抗牛脑动蛋白或动力蛋白的抗体无免疫交叉反应。最大紫外吸收波长为２７８ｎｍ，并作了ＣＤ谱分析。底物特异性研究表明：ＡＴＰａｓｅ水解活性最高。药理学研究表明：该酶蛋白可被矾酸钠强烈抑制，但对ＮＥＭ基本不敏感，氟化钠可使酶活力丧失５０％左右，寡霉素、硝酸钾及乌本苷对酶活力没有抑制作用．

Purification of the 28kD Protein from Maize Pollen and Studies on its Properties

A novel protein with ATPase activity was purified from the cytoplasmic extracts of maize pollen by acetone precipitation,ammonium sulfate fractionation,followed by DEAE-Sephadex A50 and Mono S ion-exchange chromatography.The molecular weight was about 28 kD as determined by SDS-PAGE and the isoelectric point was pH8.3 by IEF-PAGE.Western-blotting analysis showed the 28kD protein had no specific immuno-reactions with the anti-kinesin monoclonal or the anti-dynarmin polyclonal antibodies. The maximum ultraviolet absorbance was at 278nm.CD spectrum analyais showed the that 28 kD protein with the feature of a globulin.Pharmacological studies indicated that the enzyme activity was strongly inhibited by Na3VO4 but insensitive to NEM. It was inhibited about 50% by NaF. Oligomycin,KNO3 and ouabain had no effects on its ATPase activity.