王建华, 吴英杰, 吴显荣
北京农业大学生物学院.北京 100094；中科院生物物理研究所生物大分子国家重点实验室.北京 100101
Purification of 32 kD protein from maize pollen and study on some of its properties
Wang Jianhua, Wu Yingjie, Wu Xianrong
College of Biological Sciences, Beijing Agricultural University. Beijing 100094； China
A novel 32 kD protein was purified from the cytoplasmic extracts of maize pollen by the preparation of acetone powder, ammonium sulfate fractionation, followed by DEAE-Sephadex A50 and FPLC ion-exchange chromatography. The final product of this procedure showed a single band (silver staining) on the SDS-PAGE. The purification fold was 84.5. The molecular weight of its subunit was about 32 kD as determined by SDS-PAGE and the isoelectric point was pH 5.25 by IEF-PAGE. Western-blot analysis showed that the 32 kD protein did not have specific immuno-reactions with anti-kinesin and anti-dynamin polyclonal antibodies. CD spectrum analysis showed that 32 kD protein had the features of globulin. The maximum ultraviolet absorbance was at 282 nm. Its GTPase activity was higher than its ATPase activity. It was activated by Ca~(2+) and Mg~(2+). The GTPase activity of the protein was insensitive to Na_(3)VO_(4) 100 #mu#mol/L and NEM 0.1 mmol/L, indicating that it was distinct from the motor proteins such as kinesin or dynamin.
王建华, 吴英杰, 吴显荣. 玉米花粉32 kD蛋白的纯化及性质[J]. 生理学报 1996; 48 (4): .
Wang Jianhua, Wu Yingjie, Wu Xianrong. Purification of 32 kD protein from maize pollen and study on some of its properties. Acta Physiol Sin 1996; 48 (4): (in Chinese with English abstract).