两种胰蛋白酶酶解羊肌肉总蛋白样品的质谱分析

Digestion Efficacy of two Types of Trypsin on Sheep Muscles Protein via Comparative-proteomic Analysis

目的比较国产和进口两种胰蛋白酶对蛋白质的酶解效果,为后续蛋白质组学稳定研究提供依据。方法用超声破碎法提取羊肌肉组织总蛋白,经两种胰蛋白酶酶解后液相色谱串联质谱检测酶解肽段,将结果上传蛋白质序列数据库,利用质谱分析鉴定方法分析比较两种胰蛋白酶的酶解效果,并借助生物信息学工具对所得数据进行分析。结果两种胰蛋白酶酶解同种蛋白质样品产生的肽段长度86%以上均为6—18个氨基酸,适用于质谱检测。酶解后肽段产生的质量国产胰蛋白酶优于进口胰蛋白酶,国产胰蛋白酶比进口胰酶酶解可多鉴定到19%的蛋白质和34%的肽段。结论进口、国产的胰蛋白酶在酶解蛋白质方面均可用于质谱检测前蛋白质的酶解消化。但国产胰蛋白酶价格明显低于进口胰酶,更具应用优势。本研究为评价胰蛋白酶酶解效果提供了一种有效、全面的研究方法。

Objection To examine the Digestion efficacy of two types of trypsin, one is purchased domestically and the other is from foreign company, on sheep muscles protein via comparative - proteomic analysis. Method Total protein sample of sheep muscles were extracting the by sonication methods. Then, protein samples were digested into peptides by two kinds of trypsin and examined by Liquid chromatography tandem mass spectrometry. Protein sequence databases and bioinformatics tools were used to analyze mass spectrometry data. Result More than 86% of peptides from sheep muscles digested by with two kinds of trypsin were 6--18 amino acids length. More than 19% proteins or 34% peptides can be identified from sheep muscles digested by trypsin purchased domestically than that by trypsin from foreign company. Conclusion The efficacy of trypsin purchased domestically is much better than trypsin from foreign company. And the price of the domestic is lower than the imports.