不同产地太子参的iTRAQ定量蛋白质组学研究

华愉教; 王胜男; 邹立思; 刘训红; 徐建亚; 罗益远; 刘娟秀

doi:10.7538/zpxb.youxian.2016.0007

不同产地太子参的iTRAQ定量蛋白质组学研究

南京中医药大学,江苏南京210023

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- 刊出日期: 2016-05-19

iTRAQ-Based Quantitative Proteomics of Pseudostellariae Radix from Different Habitats

Nanjing University of Chinese Medicine, Nanjing 210023, China

摘要

摘要: 采用同位素标记相对和绝对定量（iTRAQ）技术对不同产地的太子参进行定量蛋白质组学研究，探讨蛋白质组表达水平的差异。所提取的太子参蛋白质样品经FASP酶解、iTRAQ试剂标记、高pH-RPLC分离、RPLC-MS分离分析，获取的串联质谱数据通过Protein Pilot 5.0软件搜库进行蛋白质鉴定，通过蛋白质相对定量的比较寻找差异表达蛋白；再对差异蛋白质进行GO（gene ontology）、KEGG代谢通路和STRING网络通路分析。实验共鉴定出3 775个蛋白质，其中3676个蛋白质具有定量信息，与传统产区相比，种植基地太子参上调差异蛋白质54个，下调差异蛋白质86个；通过生物信息学分析得到44个目标差异蛋白，主要分为9类：热休克蛋白、氧化还原酶、转移酶、水解酶、裂解酶、异构酶、Rubisco大亚基结合蛋白、伴侣蛋白质、胞腔结合蛋白。结果显示，传统产区太子参中氧化还原酶和转移酶的分解代谢、碳水化合物代谢以及抗应激能力比种植基地太子参强，但热休克蛋白、异构酶、Rubisco大亚基结合蛋白、伴侣蛋白质和胞腔结合蛋白中的蛋白质折叠和抗应激能力比种植基地太子参弱，水解酶和裂解酶的分解代谢能力与种植基地太子参相差不大；ADG1和TKTA可能是调节不同产地太子参蔗糖变化的2个关键蛋白；MFP2是导致不同产地太子参脂肪酸差异的关键蛋白。本研究可为解析不同产地太子参次生代谢物差异的成因及其药材品质形成的蛋白质机制提供参考。
- 太子参 /
- 不同产地 /
- 蛋白质组学 /
- iTRAQ
Abstract: iTRAQ proteomics approach was developed to investigate differentially expressed proteins in Pseudostellariae Radix from different habitats. The extracted proteins were digested by FASP, identified with iTRAQ coupled with LC-MS/MS and then analyzed by Protein Pilot 5.0 search engine, through the comparison of relative quantitative protein in search of differentially expressed proteins. The analysis of differentially expressed proteins was conducted using GO (gene ontology), KEGG and STRING. A total of 3 775 protein are detected, among them, 3 676 proteins are provided quantitative information, of which 54 proteins are up-regulated and 86 are down-regulated in traditional fields of Pseudostellariae Radix. 44 significantly differential expressed proteins are found, which are classified into nine categories, such as Heat shock proteins, Oxidoreductases, Transferases, Hydrolases, Lyases, Isomerases, RuBisCO large subunit-binding protein, Chaperone protein, Luminal-binding protein. The results indicated that catabolic, carbohydrate metabolism and respond to stress of Oxidoreductases and Transferases in traditional fields of Pseudostellariae Radix are stronger than other cultivated habitats of Pseudostellariae Radix, protein folding and respond to stress of Heat shock proteins, Isomerases, RuBisCO large subunit-binding protein, Chaperone protein, Luminal-binding protein are weaker, catabolic of Hydrolases and Lyases are no difference between traditional fields of Pseudostellariae Radix and other cultivated habitats. ADG1 and TKTA are the important proteins to regulate sucrose in Pseudostellariae Radix from different habitats, while MFP2 accommodate the fatty acids. This work can provide the basic information for exploring the cause of different habitats for Pseudostellariae Radix secondary metabolites and the mechanism of proteins for quality forming process.
- Pseudostellariae Radix /
- different habitats /
- proteomics /
- iTRAQ

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参考文献(27)

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不同产地太子参的iTRAQ定量蛋白质组学研究

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iTRAQ-Based Quantitative Proteomics of Pseudostellariae Radix from Different Habitats

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