化学学报 ›› 2002, Vol. 60 ›› Issue (9): 1672-1676. 上一篇    下一篇

研究论文

微透析采样荧光法研究6-硫鸟嘌呤与蛋白结合作用

王畅;吕家根;章竹君   

  1. 陕西师范大学化学与材料科学学院,西安(710062)
  • 出版日期:2002-09-15 发布日期:2002-09-15

Spectrofluorimetric Analysis Combined with Microdialysis Sampling for Studying Binding of Thioguanine to Protein

Wang Chang;Lu Jiagen;Zhang Zhujun   

  1. College of Chemistry and Materials Science, Shaanxi Normal University,Xi'an(710062)
  • Online:2002-09-15 Published:2002-09-15

将微透析技术与荧光分析相结合,测定了6-硫鸟嘌呤(Thioguanine,6-TG) 与牛血清白蛋白的相互作用。在NaOH(1.0 * 10~(-3)mol·L~(-1))介质中,6-TG可 被KMnO_4氧化,氧化产物具有较强的荧光,且荧光强度与6-TG浓度可在2.0 * 10~ (-11) ~ 2.0 * 10~(-6)mol·L~(-1)范围内分段拟合呈良好的线性关系,检测限 为4.0 * 10~(-12)mol·L~(-1)。当灌流速度为5μL·min~(-1)时,6-TG的透析率 为(7.2 ± 0.2)%。利用Scatchard方程处理数据,得到6-TG-BSA的结合常数和结合 位点分别为1.02 * 10~4 (mol·L~(-1))~(-1)和1.63。

关键词: 荧光分光光度法, 硫鸟嘌呤, 牛血清白蛋白

The binding of thioguanine (6-TG) to bovine serum albumin was studied using the technique of microdialysis combined with spectrofluorimetric analysis. In the presence of NaOH (1.0 * 10~(-3) mol·L~(-1)), 6-TG can be oxidized by KMnO_4 (5.0 * 10~(-6)mol·L~(-1)) to 2-amino-6-guanine-sulfate acid, which produces higher fluorescence emission. The fluorescence was measured at 406 nm with excitation at 329 nm. The calibration curve was linear over the concentration range of 2.0 * 10~(-11) ~ 2.0 * 10~(-6)mol·L~(-1) for 6-TG with a detection limit of 4.0 * 10~(-12)mol·L~(-1). The microdialysis probe was utilized to sample the mixed solution at a perfusion rate of 5 μL ·min~(-1) and the recovery of 6-TG under experimental condition was (7. 2 ± 0.2)%. The data obtained by the present microdialysis fluorimetric method was analyzed with the Scathard analysis plot. The estimated association constant (K) and the number of the binding sites (n) on one of BSA were 1.02 * 10~(-4) (mol·L~(-1))~(-1) and 1.63 (Scatchard equation), respectively.

Key words: FLUOROSPECTROPHOTOMETRY, thioguanine, BOVINE SERUM ALBUMIN

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