水合Ce3+离子发射强荧光, 而用Ce3+滴定植物钙调素(CaM)时, Ce3+离子荧光被完全淬灭, 据此建立了测定Ce3+离子与植物钙调素结合位点数及逐级结合常数的方法. 实验表明, Ce3+离子与植物钙调素的荧光激发光谱及发射光谱形状与光谱强度都有显著差别, 前者的荧光强度远大于后者, 故CaM存在时可以测定游离Ce3+的浓度. Ce3+离子荧光淬灭法测定结果说明, 在0.5 mol·L-1 KCl溶液中, CaM有4个Ce3+离子结合位点. 进一步通过Ce3+滴定植物钙调素的实验, 并结合Excel规划求解法, 计算出Ce3+与CaM的结合常数, K1=4.17×107, K2=1.26×107, K3=1.89×106, K4=2.04×106.
参考文献
[1] | Richdarson F S .Terbium and europium ions as lumininescent probes and stains for biomolecular systerms[J].Chemical Reviews,1982,82:541. |
[2] | Horrocks W D Jr .Luminescence spectroscopy[J].Methods in Enzymology,1993,226:495. |
[3] | Steven T F;Horrocks W D Jr .Complexation,luminescence,energy transfer of Ce3+ complexes with a series of multidentate amino phosphonic acids in aqueous solution[J].Inorganic Chemistry,1991,30:1073. |
[4] | Meng Jianxin,Gong Menglian,Tu Huamin,Yang Yansheng.Determination of Formation Constants of Ce3+ Complexes by Fluorescence[J].稀土学报(英文版),1998(02):85-89. |
[5] | 孙大业;郭艳林;马力耕;崔素娟.细胞信号转导[M].北京:科学出版社,2001:91. |
[6] | Ma, L;Cui, S;Sun, D;Xu, X .The presence of a heterotrimeric G protein and its role in signal transduction of extracellular calmodulin in pollen germination and tube growth[J].The Plant Cell,1999(7):1351-1363. |
[7] | Sun Y;Liu DL;Yu ZQ;Zhang Q;Bai J;Sun DY .An apoplastic mechanism for short-term effects of rare earth elements at lower concentrations[J].Plant, Cell and Environment,2003(6):887-896. |
[8] | 白娟;刘洪涛;孙大业 .转CaM基因工程菌的培养及重组钙调素的纯化[J].河北师范大学学报,1998,22(03):389. |
[9] | 刘德龙,孙大业,YANG Yan-Sheng,ZHANG Hong-Jie,杨燕生,张洪杰,王淑彬,龚孟濂.植物细胞外钙调素的稀土发光探针研究[J].高等学校化学学报,2000(06):860-864. |
[10] | Zhang M.;Ikura M.;Tanaka T. .CALCIUM-INDUCED CONFORMATIONAL TRANSITION REVEALED BY THE SOLUTION STRUCTURE OF APO CALMODULIN[J].Nature structural biology,1995(9):758-767. |
[11] | Cox J A .Interaction properties of calmodulin[J].Biochemical Journal,1988,249:621. |
[12] | Hui OY.;Vogel HJ. .Metal ion binding to calmodulin: NMR and fluorescence studies[J].BioMetals: An International Journal on the Role of Metal Ions in Biology, Biochemistry and Medicine,1998(3):213-222. |
上一张
下一张
上一张
下一张
计量
- 下载量()
- 访问量()
文章评分
- 您的评分:
-
10%
-
20%
-
30%
-
40%
-
50%