化学修饰辅助的蛋白质组研究策略的建立与初步应用

发布时间：2018-04-19 03:19

本文选题：磺化 + 同系物标记　；参考：《中国人民解放军军事医学科学院》2006年博士论文

【摘要】：本论文针对当前蛋白质组学研究中肽段混合物复杂、蛋白质末端信息难以获得和质谱鉴定可信度低等问题，将磺化修饰或同系物标记技术与色谱或凝胶电泳分离技术相结合，提出了蛋白质标签肽的富集、识别和鉴定的四种策略，并应用于蛋白质混合物中组氨酸肽或末端肽的选择性富集或识别，以降低体系的复杂程度和其它肽段的干扰，促进肽段的从头测序和蛋白质的鉴定。全文共分四章，介绍了四种方法：前两种方法主要针对“shotgun”技术路线，从蛋白质混合物酶切产生的肽段混合物中分离富集蛋白质的末端肽或组氨酸肽以降低体系复杂程度；后两章主要针对基于二维电泳的技术路线，通过磺化修饰或同系物标记对蛋白质的N端肽进行选择性识别。本研究首次将磺化修饰与同系物标记技术应用于蛋白质末端肽和组氨酸肽的富集、识别与序列分析，建立了四种新方法，发表了2篇中文论文，撰写了4篇英文论文，申请了一项国家自然科学基金项目，申报了四项国家发明专利。对标准蛋白质及其简单混合物的研究结果表明，本论文所建立的四种方法可以针对不同的技术路线，从蛋白质或肽段混合物中选择性富集或识别蛋白质的末端肽段或含组氨酸肽段，不仅降低了样品的复杂程度，，提高了末端肽段的丰度，而且促进了肽段的质谱测序与鉴定，提高了质谱鉴定的可信度。分析表明，这些方法在蛋白质组研究中具有应用前景。
[Abstract]:In this paper, we combine sulfonation modification or homologue labeling with chromatography or gel electrophoresis to solve the problems of complex peptide mixture in proteomics research, difficult to obtain protein terminal information and low reliability of mass spectrometry identification.Four strategies for the enrichment, recognition and identification of protein labeled peptides were proposed and applied to the selective enrichment or recognition of histidine peptides or terminal peptides in protein mixtures in order to reduce the complexity of the system and the interference of other peptide segments.To promote the ab initio sequencing of peptides and the identification of proteins.This paper is divided into four chapters. Four methods are introduced: the first two methods mainly aim at "shotgun" technology route, separating and enriching protein terminal peptide or histidine peptide from peptide segment mixture of protein mixture in order to reduce the complexity of the system;In the last two chapters, the N-terminal peptides of proteins are selectively recognized by sulfonation modification or homologue labeling.In this study, sulfonation modification and homology labeling techniques were applied to the enrichment, recognition and sequence analysis of protein terminal peptides and histidine peptides for the first time. Four new methods were established, 2 Chinese papers were published and 4 English papers were written.Applied for a National Natural Science Foundation project and declared four national invention patents.The results of the study on standard protein and its simple mixture show that the four methods developed in this paper can be used for different technical routes.The selective enrichment or recognition of the terminal peptide or histidine peptide from a protein or peptide segment mixture not only reduces the complexity of the sample, increases the abundance of the terminal peptide, but also promotes the sequencing and identification of the peptide by mass spectrometry.The reliability of mass spectrum identification is improved.The analysis shows that these methods have potential application in proteome research.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R341

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