摘要
生物分子参与生命体的整个生命活动周期,对其进行定量分析是深入研究其生物合成、信号通路和生物与非生物胁迫反应分子作用机制所关注的重要问题.尽管同位素稀释法以其在绝对定量分析结果精准方面的优势在生物分子定量分析领域得到广泛认可,但受到同位素内标化合物的限制无法得到广泛应用.近10年来同位素标记技术得到迅猛发展,尤其在定量蛋白质组、代谢组研究领域.蛋白质、多肽和其他生物小分子能够通过细胞或组织培养,或者利用化学手段,通过对特定官能团的化学反应实现同位素标记,如细胞培养氨基酸稳定同位素标记技术(SILAC),15N/14N及相对和绝对定量同位素标记技术(iTRAQ)等实现同位素标记,并结合色谱/质谱技术实现目标生物分子的定量分析.本文对上述同位素标记技术的优缺点进行了系统论述和比较.基于高效、高通量和低成本同位素标记技术的生物分子相对定量研究是未来的发展方向,也是深入了解各种生命现象分子作用机制的重要手段.
Biological molecules involve the whole life cycle of the organism. Quantitative analysis of biological molecules is a key issue for in depth understanding the molecular mechanism of these compounds, such as biosynthesis, signaling and response to biostress and abiostress. Although isotope dilution strategy is still adopted in many cases for precise quantification of target biological molecules for its overwhelming advantages, the isotope labeled internal standard set a hurdle on the way of its routine application. More and more stable isotope labeling approaches have been developed to label the target compounds efficiently during the past 10 years, especially in the field of quantitative proteomics and metabolomics. The proteins, peptides, acids and other biological molecules can be labeled by incorporated the light/heavy isotopes into organism with cell culture or growth on the isotope containing medium, or by derivatizing with isotope labeled reagents which can react with specific functional groups on biological molecules, such as SILAC, 15N/14N and iTRAQ et al, then the relative quantitative information will be obtained using chromatography/mass spectrometry （XC/MS）. The advantages and disadvantages of these isotope labeling strategies are discussed in this review. The trend of relative quantification of biological molecules based on isotope labeling is toward efficiency, high throughput and low cost in the future, which is an important method for in depth understanding the molecular mechanism of the phenomenon of life.
出处
《科学通报》
EI
CAS
CSCD
北大核心
2013年第27期2762-2778,共17页
Chinese Science Bulletin
基金
国家自然科学基金(90917017
91117016)
中国科学院仪器设备功能开发技术创新项目(YG2010002)资助
关键词
同位素标记
相对定量
质谱
生物分子
SILAC
ICAT
ITRAQ
stable isotope labeling, relative quantification, mass spectrometry, biological molecules, SILAC, ICAT, iTRAQ
