HEPES增强Tb3+和Tb2Tf的荧光发射峰强度

HEPES增强Tb3+和Tb2Tf的荧光发射峰强度

杜萍¹, 袁兰^2*, 吴后男², 徐克宁³, 杨晓改^1*

1. 北京大学药学院化学生物学系, 北京 100191
2. 北京大学医药卫生分析中心, 北京 100191
3. 北京大学天然药物及仿生药物国家重点实验室, 北京 100191

收稿日期:2009-05-05 修回日期:2009-08-10 出版日期:2009-09-15 发布日期:2009-09-15
通讯作者: 袁兰*, 杨晓改*

Enhancement of fluorescence of terbium and transferrin-bound diterbium by HEPES

Ping Du¹, Lan Yuan^2*, Hou-Nan Wu², Ke-Ning Xu³, Xiao-Gai Yang^1*

1. Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China;
2. Peking University Medical and Health Analysis Center, Peking University, Beijing 100191, China;
3. State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing 100191, China

Received:2009-05-05 Revised:2009-08-10 Online:2009-09-15 Published:2009-09-15
Contact: Lan Yuan*, Xiao-Gai Yang*

摘要/Abstract

摘要： Tb作为镧系元素中少数几个有荧光性质的稀土离子, 在荧光方面有很多重要的应用。如Tb被广泛用于与蛋白质钙位点结合的荧光探针, 同时也可根据Tb的荧光特性, 用荧光方法定量分析Tb-有机配体配合物。本研究发现HEPES (常用的pH缓冲剂) 可显著增强Tb在585 nm的特征荧光发射峰, 同时微弱的增强Tb在490 nm和549 nm的特征荧光发射峰。一直以来, 经常使用Tb在549 nm处的特征荧光发射峰定量研究Tb化合物的浓度; 然而, 本研究却发现在进行此类实验时需要慎重选择实验条件, 因为Tb荧光的定量分析结果会随缓冲试剂浓度或者含量的变化而改变。另一方面, Tb 585 nm处的特征荧光发射峰的荧光强度与HEPES和Tb离子的浓度都有一定的依赖关系, 因此Tb 585 nm特征荧光发射峰也有被用于荧光定量分析研究的潜能。

关键词: 铽离子, HEPES缓冲液, 荧光, 铽饱和转铁蛋白, 脱铁转铁蛋白

Abstract: Terbium (Tb) has been extensively used as a fluorescence probe for the identification of calcium-binding sites in proteins and for fluorometric analysis of organic ligands. In the current study, we reported that HEPES, a commonly used pH buffer reagent, significantly enhanced the characteristic emission of Tb at 585 nm. The maximum emission of Tb at 490 nm and 549 nm were also enhanced by HEPES to a less extent. Thus, cautions should be taken when quantitative analysis is performed based on the fluorescence emission of Tb at 549 nm, since the emission may vary due to the buffer reagents. Additionally, the fluorescence intensity at 585 nm was proportional to the concentration of both HEPES and terbium ions, which might be utilized to develop new fluorometric analytical methods.

Key words: Tb3+, Tb3+, HEPES, HEPES, Fluorescence, Fluorescence, Tb2Tf, Tb2Tf, ApoTf , ApoTf

中图分类号:

R916.03

Supporting:

Abbreviations: Tf, transferrin; ApoTf, apotransferrin; TfR, transferrin receptor; Tb, terbium; Tb2Tf, transferrin-bound diterbium; HEPES, N-(2-hydroxyethyl) piperazine-N'-(2-ethanesulfonic acid).
Foundation items: National Natural Science Foundation of China (Grant No. 20637010 and 20871008).
^*Corresponding author. Tel.: 86-10-82802389; e-mail: yuan_lan@bjmu.edu.cn

杜萍¹, 袁兰^2*, 吴后男², 徐克宁³, 杨晓改^1*. HEPES增强Tb3+和Tb2Tf的荧光发射峰强度[J]. .

Ping Du¹, Lan Yuan^2*, Hou-Nan Wu², Ke-Ning Xu³, Xiao-Gai Yang^1*. Enhancement of fluorescence of terbium and transferrin-bound diterbium by HEPES [J]. .

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