超滤法结合纳升液相色谱-串联质谱技术分析漏出性胸腔积液中多肽组分  

Peptidome Analysis of Transudative Pleural Effusion by Ultra-filtration Coupled with Nano-Liquid Chromatography-Tandem Mass Spectrometry

作　　者：陈世范[1] 杨静波[1] 张敏[1] 李晓鸥[2] 刘晓峰[2] 郭宏华[3] 何成彦[3] 高洪文[1] 

机构地区：[1]吉林大学第二医院,长春130041 [2]吉林省肿瘤医院,长春130021 [3]吉林大学中日联谊医院,长春130041

出　　处：《分析化学》2017年第2期224-230,共7页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.81572082;81472030);吉林省科技厅项目(Nos.20150101122JC;20150204001YY;20140311092YY)资助~~

摘　　要：漏出性胸腔积液中的多肽和蛋白质等生物分子直接或间接地与机体特定的生理、病理状态相关,反映了肺部或者全身其它部位疾病的信息。本研究利用超滤法将漏出性胸腔积液中的多肽组分进行分离,经脱盐富集后,进行纳升液相色谱-串联质谱分析。结果表明,在漏出性胸腔积液中共鉴定到来源于52种蛋白的314条多肽,超过一半的肽段来源于纤维蛋白原,并且许多肽段具有阶梯序列的特征。此外,在来源于胶原蛋白和纤维蛋白原的多肽中还发现了大量的脯氨酸氧化修饰。基因本体论富集分析显示,漏出性胸腔积液多肽组分所属蛋白均具有胞外分泌的属性。本研究给出了漏出性胸腔积液中多肽组的序列、等电点、分子量、翻译后修饰等理化参数的分布特征,为进一步寻找肺部疾病相关的多肽标志物提供了可借鉴的参考数据和分析方法。The peptides,proteins and other biological molecules in transudative pleural effusion correlate directly or indirectly with specific physiological and pathological state,reflecting the information regarding the lungs or other parts of the body.In the present study,the peptide fraction in transudative pleural effusion was isolated by ultrafiltration.After desalted and enriched by C（18） tips,the peptide mixture was analyzed by nano LC-MS/MS.The results showed that 314 peptides,which were originated from 52 proteins,in pleural transudate were identified.More than half of the peptides were derived from fibrinogen.Many peptides were characterized as displaying ladder sequences.In addition,a large number of proline oxidation modifications were detected in the peptides derived from collagen and fibrinogen.Gene ontology enrichment analysis showed that the most of the proteins extracellular properties of pleural transudate polypeptide components were protein with exocytosis.The study provided a rapid and efficient separation and analysis methods for lung disease markers related peptide compounds in pleural fluid leakage.Also this research provided a rapid and effective method for screening peptide biomarkers related to lung diseases from transudative pleural effusion.

关 键 词：多肽组 超滤 纳升液相色谱-串联质谱 漏出性胸腔积液 

分 类 号：R561.3[医药卫生—呼吸系统] O657.63[医药卫生—内科学]