摘要
羊绒与羊毛纤维蛋白质组同源性极高,这给寻找种属特异性蛋白分子标志物带来了巨大的挑战。为此,本研究提出了一套筛选、验证、定量检测羊绒分子标志物的蛋白质组研究策略。首先,采用化学标记方法,结合液相色谱串联高分辨质谱(LCMS/MS)解析羊绒、羊毛纤维蛋白分子组成,筛选羊绒、羊毛纤维候选标志物。其次,采用新型质谱平行反应监测(parallel reaction monitoring,PRM)技术,确定羊毛的4条肽段和羊绒的2条肽段作为各自的分子标志物。最终通过双甲基标记靶向定量方法结合PRM技术,绘制羊绒肽段标志物定量标准曲线,建立羊绒纤维成分定量方法。实验结果显示,研究确定的肽段分子标志物可准确鉴定混合纤维的组成成分,羊绒含量实测值与理论值的偏差<15%。所建立的发现和验证分子标志物的靶向蛋白质组研究策略,促进了更准确、更客观的新型羊绒制品质量检验方法的发展,同时可为其他高同源性蛋白质组样品中生物标志物的筛选提供借鉴。
It is very challenging to distinguish and quantify the cashmere fiber from wool fibers due to their highly similar physical appearance and substantial protein sequence homology. To address this issue, the proteomics research strategies were proposed for selecting, verifying and quantifying biomarkers for cashmere textiles. Firstly, the stable isotope labeled quantitative method and LC-MS/MS were combined to analyze proteins from cashmere and wool fibers, consequently the candidate markers were se-lected. Secondly, several specific-species biomarkers were verified using targeted MS acquisition method with parallel reaction monitoring （PRM） , altogether 4 peptides from wool and 2 peptides from cashmere were considered to be the candidate peptide markers. Finally, combining dimethyl labeling and PRM strategy, the standard cures of cashmere candidate peptide markers was established, and the quantitative method was validated with standard blended samples. As a result, the relative error between ex-perimental and theoretical is less than 15%. In conclusion, a method has been established which can discover and verify biomark-ers efficiently, and can provide the objective and robust assay for assessing authenticity of fibers and textiles. The proteomic strategies presented in this work can be further implicated in discovery of biomarkers for other samples containing highly homolo-gous proteomes, also provide technical support for commercial textile components identifying or discovering quality trait markers of other products containing highly homologous proteomes.
出处
《中国科技论文》
CAS
北大核心
2016年第6期663-669,共7页
China Sciencepaper
基金
国家自然科学基金资助项目(21505151)
天津市科技支撑计划重点项目(14ZCZDSY00062)
关键词
生物化学与分子生物学
分子标志物
双甲基标记
平行反应监测
生物质谱
biochemistry and molecular biology
biomarkers
dimethyl labeling
parallel reaction monitoring
bio-mass spectrometry
