液相層析－串聯式質譜技術於蛋白質一級結構分析的應用

Liquid Chromatography-Tandem Mass Spectrometry in Protein Primary Structure Analyses

蔡有光(Yeou-Guang Tsay)

《化學》 65卷2期 (2007/06) Pp. 105-112

https://doi.org/10.6623/chem.2007012

摘要

蛋白質一級結構包括胺基酸序列與後轉譯修飾兩部分，是蛋白質特性研究的目標之一。液相層析串聯式質譜儀(LC-MS/MS)技術，近來逐漸成為蛋白質結構分析的核心工具之一。本文將以探討蛋白質一級結構分析樣本的特性與LC-MS/MS所得資料的內涵著手，討論液相層析串聯質譜於蛋白質一級結構分析的理論基礎，作為檢討與發展分析方法的依據。並將回顧過去數年中，本實驗室於蛋白質身分鑑定、核糖體碼格變移、後轉譯修飾等分析技術中，方法開發的經驗與技術應用的結果。最後，並以「反相分子生物學」(reverse molecular biology)的概念，展望此技術在蛋白質結構分析的未來發展。

關鍵字

液相層析－串聯式質譜技術 ； 蛋白質一級結構 ； 蛋白質身份鑑定 ； 核糖體碼格變移 ； 蛋白質後轉譯修飾反向分子生物學

並列摘要

Protein primary structure is defined as the linear amino acid sequence of a polypeptide along with the modifications of the backbone and the R groups, i.e. posttranslational modifications (PTM). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been becoming one of the most important tools for analyzing these structures. In this report, we will address how LC-MS/MS has been adapted for identification of the changes in the amino acid sequences, which are regulated by various mechanisms such as ribosomal frameshifting and endoproteolytic processing. We will also update the recent development of a comprehensive posttranslational modification mapping procedure by our laboratory. Through the composition of several in-house programs, we can now map a multitude of modifications in one analysis. We have been improving the robustness of this platform such that we can pursue a grand challenge, the human PTMomics project, which aims to document the full spectrum of human protein PTMs. Examples will be provided to illustrate how bioinformatics and mass spectrometric techniques become integrated to render these protein analyses feasible.

並列關鍵字

Protein primary structure ； LC-MS/MS ； Protein identification ； Ribosomal frameshifting ； Posttranslational modification