植物细胞外钙调素的稀土发光探针研究  

Studies on Plant Extracellular Calmodulin by Lanthanide Luminescence Probe

作　　者：刘德龙[1] 孙大业[2] 杨燕生[3] 张洪杰[4] 王淑彬[4] 龚孟濂 

机构地区：[1]河北师范大学化学系,石家庄050091 [2]河北师范大学生物系,石家庄050091 [3]中山大学化学系,广州510275 [4]中国科学院长春应用化学研究所,长春130022

出　　处：《高等学校化学学报》2000年第6期860-864,共5页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金!重点项目 (批准号 :3 973 0 2 3 0 );中国科学院长春应用化学研究所稀土化学与物理开放实验室资助

摘　　要：从植物中提纯了细胞外钙调素 ( Ca M) ,并利用 NAD激酶激活作用及拮抗剂的抑制作用进行了 Ca M特性试验 .利用稀土 ( Tb3+ )发光探针测得胞外 Ca M具有 4个金属离子结合位点 .敏化 Tb3+ 激发光谱结果证明其含有 1个 Tyr残基 .胞外 Ca M( Tyr)能够向 Tb3+传能并使 Tb3+特征发光增强 .根据 Forster能量传递理论测得胞外 Ca M中 Tyr→ , 位之间距离分别为 1 .1 0 4和 1 .0 56nm,它们均小于牛脑 Ca M的相应值 .此外 ,利用敏化 Tb3+ 荧光光谱研究了 Ca M拮抗剂 W7或抗体与胞外 Ca M的相互作用 ,表明 W7或抗体优先与胞外 Ca M的 , 位半分子结合 ,使胞外 Ca M构象发生变化并导致 Tb3+ 发光强度下降 .实验表明 ,稀土发光方法不仅能获得 Ca M的结构信息 ,还可用于研究与 CaPlant extracellular calmodulin(CaM) has been purified from cauliflower and identified with NAD kinase(NADK) activation and inhibition effect of CaM antagonist W7. Tb 3+ fluorescence titration showed that extracellular CaM contained four metal binding sites. The excitation spectrum and emission specturm indicated that extracellular CaM contained one tyrosine residue which could transfer energy to bound Tb 3+ . Based on Frster type nonradiative energy transfer theory, the distances of Tyr→sites Ⅲ, Ⅳ have been determined, these are 1 104 nm(Tyr→Ⅲ, site) and 1 056 nm(Tyr→Ⅳ, site). By studing the effect of CaM antagonist W7 and CaM antibody on Tb 3+ sensitized fluorescence, it was found that the binding sites of W7 and antibody were located on the c terminal part of plant extracellular CaM which contains domain Ⅲ and domain Ⅳ. [WT5HZ]

关 键 词：细胞外钙调素 稀土发光探针 钙调素拮抗剂 植物 

分 类 号：Q942.6[生物学—植物学] O657.3[理学—分析化学]