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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 Forster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 Ca M作用靶位等复杂体系中组分的相互作用
Extracellular calmodulin (CaM) was purified from plants and tested for CaM properties using NAD kinase activation and antagonist inhibition. Extracellular CaM was measured using a rare earth (Tb3 +) luminescent probe with 4 A metal ion-binding site sensitized Tb3 + excitation spectroscopy results show that it contains a Tyr residue.External CaM (Tyr) to Tb3 + energy transfer and make Tb3 + characteristics of luminescence measured according to Forster energy transfer theory The distance between Tyr → and extracellular Ca M in extracellular Ca M was respectively 1.10 4 and 1. 566 nm, which were less than those of CaM in bovine brain.In addition, Ca M antagonist W7 or the interaction between the antibody and extracellular Ca M, indicating that W7 or antibody priority and extracellular Ca M, half of the molecular binding, the extracellular Ca M conformation change and lead to Tb3 luminescence intensity decreased .Experiments show that the rare earth luminescence method It not only can obtain the structural information of CaM, but also can be used to study the interaction of components in complex systems such as CaM target