Cell-free synthesis of stable isotope-labeled internal standards for targeted quantitative proteomics

被引：16

作者：

Narumi, Ryohei ^{[1
]}

Masuda, Keiko ^{[2
]}

Tomonaga, Takeshi ^{[1
]}

Adachi, Jun ^{[1
]}

Ueda, Hiroki R. ^{[3
,4
]}

Shimizu, Yoshihiro ^{[2
,5
]}

机构：

[1] Natl Inst Biomed Innovat Hlth & Nutr, Lab Proteome Res, 7-6-8 Satio Asagi, Ibaraki, Osaka 5670085, Japan

[2] RIKEN Quantitat Biol Ctr QBiC, Lab Single Cell Mass Spectrometry, 6-2-3 Furuedai, Suita, Osaka 5650874, Japan

[3] Univ Tokyo, Grad Sch Med, Dept Syst Pharmacol, Bunkyo Ku, 7-3-1 Hongo, Tokyo 1130033, Japan

[4] RIKEN Quantitat Biol Ctr QBiC, Lab Synthet Biol, 6-2-3 Furuedai, Suita, Osaka 5650874, Japan

[5] RIKEN Quantitat Biol Ctr QBiC, Lab Cell Free Prot Synth, 6-2-3 Furuedai, Suita, Osaka 5650874, Japan

来源：

SYNTHETIC AND SYSTEMS BIOTECHNOLOGY | 2018年 / 3卷 / 02期

基金：

日本学术振兴会;

关键词：

Absolute quantification; Mass spectrometry; Cell-free protein synthesis system; In vitro translation; Targeted quantitative proteomics; PURE system; FREE PROTEIN-SYNTHESIS; MULTIPLEXED ABSOLUTE QUANTIFICATION; MASS-SPECTROMETRY; SYNTHESIS SYSTEM; AMINO-ACIDS; PEPTIDES; SILAC; MOUSE; COLI; EFFICIENT;

D O I：

10.1016/j.synbio.2018.02.004

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

High-sensitivity mass spectrometry approaches using selected reaction monitoring (SRM) or multiple reaction monitoring (MRM) methods are powerful tools for targeted quantitative proteomics-based investigation of dynamics in specific biological systems. Both high-sensitivity detection of low-abundance proteins and their quantification using this technique employ stable isotope-labeled peptide internal standards. Currently, there are various ways for preparing standards, including chemical peptide synthesis, cellular protein expression, and cell-free protein or peptide synthesis. Cell-free protein synthesis (CFPS) or in vitro translation (IVT) systems in particular provide high-throughput and low-cost preparation methods, and various cell types and reconstituted forms are now commercially available. Herein, we review the use of such systems for precise and reliable protein quantification. (c) 2018 The Authors. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co.

引用

页码：97 / 104

页数：8

共 80 条

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