Protein Absolute Quantification Using Cell-Free-Synthesized Peptide

• Narumi Ryohei

  Laboratory of Proteome Research, National Institutes of Biomedical Innovation, Health and Nutrition Laboratory for Synthetic Biology, RIKEN Quantitative Biology Center

• Shimizu Yoshihiro

  Laboratory for Cell-Free Protein Synthesis, RIKEN Quantitative Biology Center Laboratory for Single Cell Mass Spectrometry, RIKEN Quantitative Biology Center

• R. Ueda Hiroki

  Laboratory for Synthetic Biology, RIKEN Quantitative Biology Center Department of Systems Pharmacology Graduate School of Medicine, The University of Tokyo

Bibliographic Information

Other Title

• 無細胞合成ペプチドを利用したタンパク質絶対定量

Abstract

Accurate measurements of absolute protein abundance can be achieved by emerging targeted proteomics approaches, in which target peptides of enzymatic digests of proteins are analyzed by selected reaction monitoring (SRM) using the stable isotope labeled peptides as internal standard. Thus, development of a simple and easy way for the preparation of internal standards is advantageous for the analyses of multiple target proteins. We developed a method, termed MS-based Quantification By isotope labeled Cell-free products (MS-QBiC), which provided the simple and high-throughput preparation of internal standards by using a reconstituted cell-free protein synthesis system called PURE system. This method facilitates both multiplexed and sensitive quantification of absolute amounts of target proteins, and then can enhance the capability of the targeted proteomics.

Journal

• Proteome Letters

  Proteome Letters 2 (2), 75-82, 2017

  Japanese Proteomics Society

Keywords

• absolute quantification
• cell-free protein synthesis system
• mass spectrometry
• selected reaction monitoring
• targeted proteomics

Details

• CRID: 1390001206229626624
• NII Article ID: 130006743386
• DOI: 10.14889/jpros.2.2_75
• ISSN: 24322776
• Text Lang: ja
• Data Source:
  • JaLC
  • CiNii Articles
• Abstract License Flag: Disallowed