建立了黄酮与溶菌酶相互作用研究的强度衰减-基质辅助激光解吸离子化-质谱(Intensity fading matrix-assisted laser desorption/ionization mass spectrometry,IF-MALDI-MS)分析方法。在优化的基质DHB条件下,分别研究了木犀草素、染料木素、芹菜素、槲皮素和大豆黄素与溶菌酶相互作用,比较了溶菌酶加入前后黄酮的相对丰度的变化,并通过竞争实验的方法研究了5种黄酮与溶菌酶结合的亲和性大小。结果表明,5种黄酮与溶菌酶均存在相互作用,亲和性强弱为:木犀草素>芹菜素,染料木素>槲皮素>大豆黄素。结合5种黄酮的结构特征,讨论了黄酮的结构及其与溶菌酶亲和性的关系,发现C5位和C3'位的羟基有利于黄酮与溶菌酶的结合,C3位的羟基不利于黄酮与溶菌酶的结合。本方法具有简便,快速,高效等优点,也可以应用于其它天然产物与蛋白质的相互作用的研究。
【英文摘要】
Flavonoids are reported to have a variety of biological activities. Investigations on the interactions of flavonoids with protein are essential to understand the biological activities of flavnoids. In this paper,by using the optimized matrix 2,5-dihydroxybenzoic acid( DHB), we investigated the interaction of five flavonoids( luteolin,apigenin,genisten,quercetin,dazein) with lyszoyme using intensity fading( IF)-MALDI-MS method. The changes of relative abundances of flavonoids before and after addition of lys...