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

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

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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 0ng·mL-1之间 )和较好的选择性 。

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), alpha-chymotrypsin (alpha-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 (sigma = 3) in the range of 4.0 similar to 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.