大连化物所为蛋白质组学研究提供新方法

佚名

   生物谷报道：美国化学会刊物——《蛋白质组学研究》(Journal  of  Proteome  Research)近日发表了中科院大连化物所研究员邹汉法与上海国家组织工程中心专家崔磊合作的关于骨组织蛋白质提取方法和蛋白质组学分析的学术论文，论文被该刊选为Research  Profile  Paper进行评述和介绍。美国化学会出版的学术刊物在每期发表的学术论文中优选若干篇重要论文进行点评。《蛋白质组学研究》作为美国化学会出版的蛋白质组学研究领域的权威性刊物之一，在每期发表的40到50篇学术论文中只选取2篇论文作为Research  Profile  Paper进行点评。

    骨组织是深度矿化的组织，有效提取骨组织中的蛋白质并进行大规模鉴定是十分困难的工作，邹汉法等人在对骨组织去矿化处理后，采用四种不同的溶剂提取骨组织中的蛋白质，通过Shotgun方法进行蛋白质组学分析，共鉴定了2479个蛋白质，其中由二段肽以上鉴定的高可信度蛋白质816个，大大超过2007年国外学者报导的鉴定133个蛋白质(Schreiweis  et  al，J.  Cell  Biochem.  2007，101，466-476)，是骨组织中鉴定蛋白质的最好结果，为骨疾病诊断标记的发现和生物学过程的研究提供了有效的方法和技术。

    邹汉法近年来广泛开展了蛋白质组学和多肽组学新技术新方法的研究，近两年来相关研究成果已在Mol.  Cel.  Proteomics(IF  9.876)，Angew.  Chem.  Int.  Ed.(IF  9.596)，Nucleic  Acids  Research(IF  7.552)，J.  Proteome  Res.(IF  6.901)，Proteomics(IF  6.088)，Anal.  Chem.(IF  5.635)等学术刊物发表学术论文15篇，申请国家发明专利5项，正在申请美国发明专利1项。相关研究成果已引起国内外的广泛关注，应邀已在Trends  Anal.  Chem.  刊物发表蛋白质组学新技术和新方法的综述论文1篇，并受邀为Proteomics刊物撰写蛋白质组学样品制备新技术和新方法的综述论文。（引自大连化学物理研究所）

原始出处:
J. Proteome Res., 6 (6), 2287 -2294, 2007. 10.1021/pr070056t S1535-3893(07)00056-5
Web Release Date: May 8, 2007 Copyright © 2007 American Chemical Society

Method Development of Efficient Protein Extraction in Bone Tissue for Proteome Analysis

Xiaogang Jiang, Mingliang Ye, Xinning Jiang, Guangpeng Liu, Shun Feng, Lei Cui,* and Hanfa Zou*

National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China, Shanghai Tissue Engineering Research and Development Center, Shanghai 200235, China, and School of Medicine, Suzhou University, Suzhou 215007, China

Received January 30, 2007

Abstract:

Exploring bone proteome is an important and challenging task for understanding the mechanisms of physiological/pathological process of bone tissue. However, classical methods of protein extraction for soft tissues and cells are not applicable for bone tissue. Therefore, method development of efficient protein extraction is critical for bone proteome analysis. We found in this study that the protein extraction efficiency was improved significantly when bone tissue was demineralized by hydrochloric acid (HCl). A sequential protein extraction method was developed for large-scale proteome analysis of bone tissue. The bone tissue was first demineralized by HCl solution and then extracted using three different lysis buffers. As large amounts of acid soluble proteins also presented in the HCl solution, besides collection of proteins in the extracted lysis buffers, the proteins in the demineralized HCl solution were also collected for proteome analysis. Automated 2D-LC-MS/MS analysis of the collected protein fractions resulted in the identification of 6202 unique peptides which matched 2479 unique proteins. The identified proteins revealed a broad diversity in the protein identity and function. More than 40 bone-specific proteins and 15 potential protein biomarkers previously reported were observed in this study. It was demonstrated that the developed extraction method of proteins in bone tissue, which was also the first large-scale proteomic study of bone, was very efficient for comprehensive analysis of bone proteome and might be helpful for clarifying the mechanisms of bone diseases.

Keywords: protein extraction bone proteome shotgun proteomics tandem mass spectrometry bone diseases biomarker discovery