高等学校化学学报

高等学校化学学报 ›› 2000, Vol. 21 ›› Issue (6): 860.

• 论文 • 上一篇    下一篇

植物细胞外钙调素的稀土发光探针研究

刘德龙1, 孙大业2, 杨燕生3, 张洪杰4, 王淑彬4, 龚孟濂4   

  1. 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
  • 收稿日期:1999-06-11 出版日期:2000-06-24 发布日期:2000-06-24
  • 通讯作者: 刘德龙(1964年出生),男,博士,副教授,从事稀土生物无机化学与分析化学研究.
  • 基金资助:

    国家自然科学基金重点项目(批准号:39730230);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

Studies on Plant Extracellular Calmodulin by Lanthanide Luminescence Probe

LIU De-Long1, SUN Da-Ye2, YANG Yan-Sheng3, ZHANG Hong-Jie4, WANG Shu-Bin4, GONG Meng-Lian4   

  1. 1. Department of Chemistry, Hebei Normal University, Shijiazhuang 050091, China;
    2. Department of Biology, Hebei Normal University, Shijiazhuang 050091, China;
    3. Department of Chemistry, Zhongshan University, Guangzhou 510275, China;
    4. Laboratoryof Rare Earth Chemistryand Physics, Changchun Instituteof Applied Chemistry, Chinese Academy of Science, Changchun 130022, China
  • Received:1999-06-11 Online:2000-06-24 Published:2000-06-24

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被引次数

37

摘要： 从植物中提纯了细胞外钙调素(CaM),并利用NAD激酶激活作用及拮抗剂的抑制作用进行了CaM特性试验.利用稀土(Tb3+)发光探针测得胞外CaM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CaM(Tyr)能够向Tb3+传能并使Tb3+特征发光增强.根据Förster能量传递理论测得胞外CaM中Tyr→,位之间距离分别为1.104和1.056nm,它们均小于牛脑CaM的相应值.此外,利用敏化Tb3+荧光光谱研究了CaM拮抗剂W7或抗体与胞外CaM的相互作用,表明W7或抗体优先与胞外CaM的,位半分子结合,使胞外CaM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CaM的结构信息,还可用于研究与CaM作用靶位等复杂体系中组分的相互作用.

关键词: 细胞外钙调素, 稀土发光探针, Tb3+, 钙调素拮抗剂

Abstract: Plant extracellular calmodulin(CaM) has been purified from cauliflower and identified with NADkinase(NADK) activation and inhibition effect of CaMantagonist W7. Tb3+ fluorescence titration showed that extracellular CaMcontained four metal-binding sites. The excitation spectrum and emission specturm indicated that extracellular CaMcontained one tyrosine residue which could transfer energy to bound Tb3+ . Based on Förster 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 CaMantagonist W7 and CaMantibody on Tb3+ sensitized fluorescence, it was found that the binding sites of W7 and antibody were located on the c terminal part of plant extracellular CaMwhich contains domain Ⅲ and domain Ⅳ.

Key words: Extracellular calmodulin, Lanthanide luminescence probe, Terbium(Ⅲ), CaM antagonist

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