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出 处:《Acta Botanica Sinica》2001年第11期1140-1145,共6页Acta Botanica Sinica(植物学报:英文版)
基 金:SupportedbytheStateKeyBasicResearchandDevelopmentPlanofChina (G19990 1170 5 ) ;theNationalNaturalScienceFoundationofChina ( 39770 0 77)andtheScienceandTechnologyProgramoftheMinistryofEducationofChina ( 99133) .
摘 要:The stimulatory effect of lysophosphatidylcholine (lyso_PC) on ATP and ρ_nitrophenyl phosphate (PNPP) hydrolysis by the plasma membrane H +_ATPase from soybean (Glycine max (L.) Merr.) hypocotyls was studied. Results showed that lyso_PC stimulated the hydrolysis of ATP; ATP hydrolysis was enhanced dramatically when lyso_PC was within 0-0.03%, and increased slightly when lyso_PC was higher than 0.03%. At the concentration of 0.03%, lyso_PC stimulated ATP hydrolysis by 80.5%. Kinetics analysis showed that V max increased from 0.46 μmol P i·mg -1 protein·min -1 to 0.87 μmol P i·mg -1 protein·min -1 while K m increased from 0.88 mmol/L to 1.15 mmol/L under lyso_PC treatment. The optimum pH of ATP hydrolysis was shifted from 6.5 to 7.0 . Moreover, it was found lyso_PC enhanced the inhibition of ATP hydrolysis by hydroxylamine. In the presence of 200 mmol/L hydroxylamine, ATP hydrolysis was inhibited by 74.4%, while it was inhibited by 84.4% when treated with lyso_PC. However, PNPP hydrolysis and the inhibitory effect of vanadate were not affected by lyso_PC. The above results indicated that the kinase domain might be an action site or regulatory region of the C_terminal autoinhibitory domain in the plant plasma membrane H +_ATPase.研究了溶血卵磷脂 (lysophosphatidylcholine,lyso PC)对大豆 (Glycinemax (L .)Merr.)下胚轴质膜H+ _ATPaseATP和对_硝基苯磷酸 (ρ_nitrophenylphosphate ,PNPP)水解的影响。结果显示 ,lyso_PC可以刺激ATP水解活力 ,当lyso_PC浓度在 0~ 0 .0 3%范围时 ,ATP水解活力明显提高 ,lyso_PC浓度高于 0 .0 3%后增加缓慢 ;当lyso_PC浓度为0 .0 3%时 ,ATP水解活力提高 80 .5 %。动力学分析表明 ,lyso_PC处理可以使ATP水解的Vmax从 0 .46 μmolPi·mg-1protein·min-1升高到 0 .87μmolPi·mg-1protein·min-1;使Km 从 0 .88mmol/L升高到 1.15mmol/L。最适pH也从 6 .5变为 7.0。实验还发现lyso_PC可以促进羟胺对ATP水解的抑制作用 ,lyso_PC处理后ATP水解被羟胺 (2 0 0mmol/L)抑制 84.4% ,而未经lyso_PC处理的对照组则仅被抑制 74.4%。然而 ,lyso_PC处理不影响PNPP水解活力和钒酸钠抑制效应。以上结果表明 ,质膜H+
关 键 词:lysophosphatidylcholine (lyso_PC) soybean hypocotyls plasma membrane H +_ATPase C_terminal autoinhibitory domain kinase domain
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