摘要
建立了一种基于螯合稀土金属标记蛋白质的技术,优化了影响标记效率的各种条件。在50mmol/LTEAB缓冲体系(pH7.5),DTPA与肽段摩尔比为100∶1,常温反应30min的条件下,DTPA标记完全;在100mmol/L乙酸铵缓冲液(pH6.0),稀土金属与DTPA摩尔比为4∶1,37℃反应1h的条件下,稀土金属螯合反应完全。考察了不同稀土金属的螯合效率,发现离子半径最小的Lu螯合效率最高。本方法被成功应用于肽混合物标记。
A strategy for protein labeling based on rare earth metal chelating as an elemental tag was established.Several influencing factors were examined to ensure the completeness and efficiency of the reaction.The optimized conditions were as follows：The first step of acylation reaction was reacted in 50 mmol/L pH 7.5 triethylammonium bicarbonate （TEAB） buffer for 30 min at room temperature,and the molar ratio of diethyl-ammionium-N,N,N＇N″,N-pentaacetie acid dianhydride （DTPA） with peptide was 100; The second step of rare earth metal chelating was in 100 mmol/L pH6 ammonium acetate buffer for 1 h at 37 ℃,and the molar ratio of rare earth metals with DTPA was 4.The chelating efficiency of different rare earth metal was compared.The resutls indicated that the metal chelating of Lu with the minimum ionic radius had the maximum efficiency.The labeling strategy was also successfully applied in peptide mixture.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2010年第10期1393-1399,共7页
Chinese Journal of Analytical Chemistry
基金
国家高技术研究发展计划(No.2006AA02A308)
国家重点基础研究发展计划(No.2006CB910801)
国家自然科学基金(No.20735005)
国家科技重大专项(No.2008ZX08012-003)资助项目
关键词
质谱
螯合稀土金属标记
蛋白质定量
Mass spectrometry
Rare-earth metal chelated tags labeling
Protein quantification
作者简介
E-mail：qianxhl@ yahoo. com. cn
