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质谱分析法在蛋白质组学中的应用。

Application of mass spectrometry in proteomics.

作者信息

Guerrera Ida Chiara, Kleiner Oliver

机构信息

Department of Medicine, Centre for Molecular Medicine, University College London, UK.

出版信息

Biosci Rep. 2005 Feb-Apr;25(1-2):71-93. doi: 10.1007/s10540-005-2849-x.

Abstract

Mass spectrometry has arguably become the core technology in proteomics. The application of mass spectrometry based techniques for the qualitative and quantitative analysis of global proteome samples derived from complex mixtures has had a big impact in the understanding of cellular function. Here, we give a brief introduction to principles of mass spectrometry and instrumentation currently used in proteomics experiments. In addition, recent developments in the application of mass spectrometry in proteomics are summarised. Strategies allowing high-throughput identification of proteins from highly complex mixtures include accurate mass measurement of peptides derived from total proteome digests and multidimensional peptide separations coupled with mass spectrometry. Mass spectrometric analysis of intact proteins permits the characterisation of protein isoforms. Recent developments in stable isotope labelling techniques and chemical tagging allow the mass spectrometry based differential display and quantitation of proteins, and newly established affinity procedures enable the targeted characterisation of post-translationally modified proteins. Finally, advances in mass spectrometric imaging allow the gathering of specific information on the local molecular composition, relative abundance and spatial distribution of peptides and proteins in thin tissue sections.

摘要

质谱法可以说是蛋白质组学的核心技术。基于质谱的技术在对源自复杂混合物的全球蛋白质组样品进行定性和定量分析方面的应用,对细胞功能的理解产生了重大影响。在此,我们简要介绍蛋白质组学实验中目前使用的质谱原理和仪器。此外,还总结了质谱在蛋白质组学应用中的最新进展。从高度复杂的混合物中高通量鉴定蛋白质的策略包括对源自全蛋白质组消化产物的肽段进行精确质量测量以及与质谱联用的多维肽段分离。完整蛋白质的质谱分析可实现蛋白质异构体的表征。稳定同位素标记技术和化学标记的最新进展使得基于质谱的蛋白质差异显示和定量成为可能,新建立的亲和方法能够对翻译后修饰的蛋白质进行靶向表征。最后,质谱成像技术的进展使得在薄组织切片中收集有关肽段和蛋白质的局部分子组成、相对丰度和空间分布的特定信息成为可能。

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