生物谷報道： 美國化學(xué)會刊物——《蛋白質(zhì)組學(xué)研究》(Journal of Proteome Research)近日發(fā)表了中科院大連化物所研究員鄒漢法與上海國家組織工程中心專家崔磊合作的關(guān)于骨組織蛋白質(zhì)提取方法和蛋白質(zhì)組學(xué)分析的學(xué)術(shù)論文,，論文被該刊選為Research Profile Paper進(jìn)行評述和介紹。美國化學(xué)會出版的學(xué)術(shù)刊物在每期發(fā)表的學(xué)術(shù)論文中優(yōu)選若干篇重要論文進(jìn)行點評,?！兜鞍踪|(zhì)組學(xué)研究》作為美國化學(xué)會出版的蛋白質(zhì)組學(xué)研究領(lǐng)域的權(quán)威性刊物之一，在每期發(fā)表的40到50篇學(xué)術(shù)論文中只選取2篇論文作為Research Profile Paper進(jìn)行點評,。
骨組織是深度礦化的組織,，有效提取骨組織中的蛋白質(zhì)并進(jìn)行大規(guī)模鑒定是十分困難的工作，鄒漢法等人在對骨組織去礦化處理后,，采用四種不同的溶劑提取骨組織中的蛋白質(zhì),，通過Shotgun方法進(jìn)行蛋白質(zhì)組學(xué)分析，共鑒定了2479個蛋白質(zhì),，其中由二段肽以上鑒定的高可信度蛋白質(zhì)816個,，大大超過2007年國外學(xué)者報導(dǎo)的鑒定133個蛋白質(zhì)(Schreiweis et al，J. Cell Biochem. 2007,，101,，466-476)，是骨組織中鑒定蛋白質(zhì)的最好結(jié)果,，為骨疾病診斷標(biāo)記的發(fā)現(xiàn)和生物學(xué)過程的研究提供了有效的方法和技術(shù),。
鄒漢法近年來廣泛開展了蛋白質(zhì)組學(xué)和多肽組學(xué)新技術(shù)新方法的研究，近兩年來相關(guān)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)等學(xué)術(shù)刊物發(fā)表學(xué)術(shù)論文15篇,，申請國家發(fā)明專利5項，正在申請美國發(fā)明專利1項,。相關(guān)研究成果已引起國內(nèi)外的廣泛關(guān)注,，應(yīng)邀已在Trends Anal. Chem. 刊物發(fā)表蛋白質(zhì)組學(xué)新技術(shù)和新方法的綜述論文1篇，并受邀為Proteomics刊物撰寫蛋白質(zhì)組學(xué)樣品制備新技術(shù)和新方法的綜述論文,。（引自大連化學(xué)物理研究所）
原始出處:
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
