蚕豆叶片下表皮ABA结合蛋白的分离纯化_吴忠义.pdfVIP

1999, 41 ( 8) : 842~ 845 A cta Botanica Sinica * ABA ** 吴忠义 张大鹏 贾文锁 ( 100094) ABA 10 ( 6~ 8 mmol/ L ) 4B , , ( Viciaf aba L. ) ABA ( ABA_BP) , NaCl , 1 mmol/ L ABA ABA_BP, ABA , 112 ABA 58. 33 nmol/ g pro ein, Kd 21 nmol/ L, 44 . 2 kD, 90 % ABA , 280 nm ABA, , , Isolation and Purification of ABA Binding Protein from Abaxial * Epiderm of Viciaf aba Leaf ** WU Zhong_Yi ZHANG Da_Peng JIA Wen_Suo ( College of Plant Science and Technology, China Agricultural University, Beijing 100094) Abstract Abscisic acid (ABA) was efficien ly cross_linked o Sepharose 4B ( 6~ 8 mmol ABA/ L gel) by an arm of 10_a om carbon chain. Solubilized ABA_BP (ABA binding pro ein) was allowed o bind o he gel, while unrela ed pro eins were removed by washing wi h a gradien of NaCl buffer. The ABA_BP was elu ed wi h 1 mmol/ L ABA . Since ABA a high concen ra ion can in erfere wi h bo h he binding ac ivi y assay and pro ein analysis, he frac ions elu ed wi h ABA were passed hrough a Sephadex G_25 column o remove he ABA. Frac ions con aining he binding ac ivi y were pooled, concen ra ed wi h ul ra_fil ra ion . The maximum binding capaci y ( B ) of he purified ABA_BP was 58. 33 nmol/ g pro ein, and he K was 21 nmol/ L, wi h an ap- MAX d proxima ely 112 fold