生物活性肽蛋白质组分分离与鉴定方法的建立

目的:评估大规模蛋白质组分离与鉴定技术策略在生物活性肽研究中的应用价值。方法:采用全溶液酶解及胶内分离与酶解方法分离生物活性肽;肽段离子阱串联质谱鉴定时,采用碰撞诱导解离与电子转移解离2种互补的肽段碎裂模式。结果:几种方法联用,鉴定到复杂生物活性肽样品中236个多肽大分子;不同分离和鉴定策略显示出良好的互补性,基于凝胶电泳分离的策略提供了最好的鉴定效果。结论:大规模的蛋白质组学分离与鉴定技术策略可以有效应用于生物活性肽组分的表征。

Establishment of Methods for Large Scale Identification of Components in Bioactive Peptide Mixture

Objective： To evaluate the applicability of large scale proteomic technologies in profiling the compo- nents in bioactive peptide mixture. Methods： In-solution or in-gel digestion were used for protein digestion, and the two ion fragmentation mode, eollisional induced-dissociation（CID） and electron transfer dissoeiation（ETD） in ion trap mass spectrometry, were used to sequence the peptides. Results： By combing different separation and identification strategy, we profiled 236 polypeptides fractions in the bioactive peptide mixture. These methods showed typical complementation, while the gel-based strategy provided the best results. Conclusion： The large scale proteomics strategy could be implemented in the effective characterization of hioactive peptide mixture.