High-throughput synthesis of stable isotope-labeled transmembrane proteins for targeted transmembrane proteomics using a wheat germ cell-free protein synthesis system

Nobuaki Takemori; Ayako Takemori; Kazuhiro Matsuoka; Ryo Morishita; Natsuki Matsushita; Masato Aoshima; Hiroyuki Takeda; Tatsuya Sawasaki; Yaeta Endo; Shigeki Higashiyama

doi:10.1039/C4MB00556B

High-throughput synthesis of stable isotope-labeled transmembrane proteins for targeted transmembrane proteomics using a wheat germ cell-free protein synthesis system†

Nobuaki Takemori,*^a Ayako Takemori,^a Kazuhiro Matsuoka,^ab Ryo Morishita,^c Natsuki Matsushita,^d Masato Aoshima,^e Hiroyuki Takeda,^ab Tatsuya Sawasaki,^ab Yaeta Endo^b and Shigeki Higashiyama^a

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* Corresponding authors

^a Proteo-Science Center, Ehime University, Ehime, Japan
E-mail: takemori@m.ehime-u.ac.jp

^b Cell-Free Science and Technology Research Center, Ehime University, Ehime, Japan

^c CellFree Sciences Co., Ltd., Ehime, Japan

^d Translational Research Center, Ehime University Hospital, Ehime, Japan

^e K.K. AB SCIEX, Tokyo, Japan

Abstract

Using a wheat germ cell-free protein synthesis system, we developed a high-throughput method for the synthesis of stable isotope-labeled full-length transmembrane proteins as proteoliposomes to mimic the in vivo environment, and we successfully constructed an internal standard library for targeted transmembrane proteomics by using mass spectrometry.

Molecular BioSystems

High-throughput synthesis of stable isotope-labeled transmembrane proteins for targeted transmembrane proteomics using a wheat germ cell-free protein synthesis system†

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High-throughput synthesis of stable isotope-labeled transmembrane proteins for targeted transmembrane proteomics using a wheat germ cell-free protein synthesis system

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