化学学报 ›› 2004, Vol. 62 ›› Issue (17): 1635-1640. 上一篇    下一篇

研究论文

某些氨羧络合型染料与蛋白质相互作用的共振瑞利散射研究

刘绍璞, 范莉, 胡小莉, 刘忠芳, 陈粤华   

  1. 西南师范大学化学化工学院, 重庆, 400715
  • 投稿日期:2004-01-05 修回日期:2004-03-22 发布日期:2014-02-17
  • 通讯作者: liusp@swnu.edu.cn
  • 作者简介:刘绍璞E-mail:liusp@swnu.edu.cn
  • 基金资助:
    国家自然科学基金(No.20175018)资助项目.

Study on the Interaction between Proteins and Some Complexon Dyes by Resonance Rayleigh Scattering Method

LIU Shao-Pu, FAN Li, HU Xiao-Li, LIU Zong-Fang, CHEN Yue-Hua   

  1. School of Chemistry and Chemical Engineering, Institute of Environmental Chemistry, Southwest China Normal University, Chongqing 400715
  • Received:2004-01-05 Revised:2004-03-22 Published:2014-02-17

用共振瑞利散射(RRS)光谱法研究了酚酞络合剂、百里酚酞络合剂以及钙黄绿素等氨羧络合型染料与人血清白蛋白(HSA)、牛血清白蛋白(BSA)、α-糜蛋白(α-Chy)、卵白蛋白(Ova)、溶菌酶(Lys)等水溶性蛋白质的相互作用,发现只有作为氨羧络合剂咕吨染料的钙黄绿素与蛋白质反应能使RRS显著增强,并观察到特征的RRS光谱,而作为氨羧络合型三苯甲烷的酚酞络合剂、百里酚酞络合剂作用不明显.文中对于钙黄绿素与蛋白质的反应条件、影响因素、反应产物的光谱特征及有关分析化学性质进行了研究,并拟定了用钙黄绿素RRS测定水溶性蛋白质的方法.该方法有高的灵敏度(对不同蛋白质的检出限在4.0~40.0 ng·mL-1之间)和较好的选择性,可用于人血清中蛋白质的测定.

关键词: 蛋白质, 共振瑞利散射, 氨羧络合型染料, 钙黄绿素

In this paper it is reported the results of interaction of complexon dyes such as phenolphthalein, thymolphthalexone and calcein with water-soluble proteins such as human serum albumin (HSA), bovine serum albumin (BSA), α-chymotrypsin (α-Chy), ovalbumin (Ova) and lysozyme (Lys).From our experiments it was discovered that only the reaction of proteins with calcein, which is a kind of complexon xanthene dye, can result in a significant enhancement of intensity of the resonance Rayleigh scattering (RRS), and the corresponding characteristic RRS spectrum can also be observed.But no obviously similar results were obtained in the similar experiments of phenolphthalein and thymolphthalexone which are complexon triphenylmethane.In this work, the RRS spectral characteristics of the combination products of calcein with protein, the optimum condition of the reactions, the primary influencing factors as well as the relevant analytical properties have been investigated systematically.Based on these research results, the resonance Rayleigh scattering method for the determination of water-soluble protein with calcein was established.The new RRS method has high sensitivity with the detection limits (σ=3) in the range of 4.0~40.0 ng·mL-1 for different proteins and fairly good selectivity, and can be applied to the direct determination of proteins in human serum.

Key words: protein, resonance Rayleigh scattering, complexon dye, calcein