摘要
通过酸酐同系物标记蛋白质的N-末端,经过凝胶电泳分离,使用基质辅助激光解析电离飞行时间质谱仪进行鉴定,建立了一种基于化学标记和质谱技术的、用于蛋白质的N-末端鉴定和相对定量新方法。标记后的蛋白质的N端肽以相差14 Da的成对峰形式在质谱中出现,而非N-末端肽则是以单峰呈现,从而实现了末端肽的特异性识别;通过端肽的信号强度对3种标记蛋白质的不同比例的混合溶液实现了相对定量分析。
A new method was developed for quantifying proteins and identifying their N-termini using homolog labeling coupled with gel-based separation and MALDI-TOF mass spectrometry. Peak pairs with a mass difference of 14 Da were identified as N-terminal peptides while the internal peptides were single peaks and the ratios of their peak intensity were calculated for relative quantification of proteins. The results obtained from 3-protein mixtures demonstrate that this homolog labeling method enables the identification of N-terminal fragment ions from other fragments and quantitation of proteins simultaneously.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2008年第12期1606-1613,共8页
Chinese Journal of Analytical Chemistry
基金
国家重点基础研究规划项目(Nos.2006CB910803、2007CB914104)
国家高技术研究发展规划(No.2006AA02A308)
国家自然科学基金重点项目和国家自然科学基金(Nos.20635010,20735005,20505018)资助
关键词
同系物标记
化学修饰
电泳分离
基质辅助激光解析电离飞行时间质谱
N-末端肽
蛋白质组学
Homolog labeling, chemical modification, gel-based separation, matrix assisted laser desorption ionization-time of flight-mass spectrometry, N-terminal peptides, proteomics
