| 林晨,温成平,范永升.蕲蛇乙醇提物中多肽成分的蛋白组学研究[J].浙江中西医结合杂志,2017,27(5): |
| 蕲蛇乙醇提物中多肽成分的蛋白组学研究 |
| Proteomatic analysis of peptides from QiShe(Agkistrodon acutus(Guenther)) |
| 投稿时间：2016-10-25 修订日期：2017-02-20 |
| DOI： |
| 中文关键词: 蕲蛇 多肽 蛋白质组学 凝胶电泳 质谱鉴定 |
| 英文关键词:Agkistrodon acutus polypeptides proteomics electrophoresis mass spectrometry |
| 基金项目:浙江省教育厅科研项目(Y201430952)，浙江中医药大学校级科研基金人才项目(2013ZR01) |
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| 中文摘要: |
| 【目的】通过蛋白质组学方法分析蕲蛇乙醇提取物中的多肽类成分。【方法】双向凝胶电泳技术分离多肽,染色后切取多肽斑点胶内酶切,利用基质辅助激光解吸离子飞行时间串联质谱(MALDI-TOF-MS/MS)分析酶切后抽提的肽段,获取的质谱数据进一步用于数据库检索,鉴定相关肽段序列。【结果】采用稀乙醇热浸法每克蕲蛇药材多肽溶出率为5.48%。凝胶图像显示各等电点区间均有2kDa的多肽分布,无法通过串联质谱数据结合数据库检索方式有效鉴定,而在15kDa区间存在的胶点中高置信度匹配到多个肽段。【结论】采用稀乙醇热浸法能够有效提取蕲蛇干燥药材中的多肽成分。 |
| 英文摘要: |
| Purpose To carry out a proteomic analysis of peptides contained within the ethanol extracts of chinese medicine QiShe. Methods ethanol extracts were produced following steps recorded in the Chinese pharmacopeia, and were used as samples by further processing of precipitation and lyophilization. Polypeptides within the samples were initially separated by two-dimensional electrophoresis. Valid spots were cut and trypsin-proteolysed, and were analyzed using TOF-TOF mass spectrometry, data from which were search in the database to establish the identification. Results Extraction rate was 5.48% per gram crude medicine. Polypeptides separated by electrophoresis mainly distributed at 2kDa mark area, indicating their composition of 15-20 amino acids. Produced mass data from two gel spots within 10-15kDa mark area were identified in the database as fragments of skeletal alpha (α)-actin and Type I collagen. Conclusions Heating of ethanol is effective to produce extracts containing polypeptides from QiShe. Certain fragments sequences identified implied their possible roles involved in oral tolerance. |
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