Selected Reaction Monitoring for the Quantification of Escherichia Coli Ribosomal Proteins

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2020 Dec 14

PMID 33315868

Citations 1

Authors

Yuishin Kosaka

Wataru Aoki

Megumi Mori

Shunsuke Aburaya

Yuta Ohtani

Hiroyoshi Minakuchi

Mitsuyoshi Ueda

Affiliations

Soon will be listed here.

Abstract

Ribosomes are the sophisticated machinery that is responsible for protein synthesis in a cell. Recently, quantitative mass spectrometry (qMS) have been successfully applied for understanding the dynamics of protein complexes. Here, we developed a highly specific and reproducible method to quantify all ribosomal proteins (r-proteins) by combining selected reaction monitoring (SRM) and isotope labeling. We optimized the SRM methods using purified ribosomes and Escherichia coli lysates and verified this approach as detecting 41 of the 54 r-proteins separately synthesized in E. coli S30 extracts. The SRM methods will enable us to utilize qMS as a highly specific analytical tool in the research of E. coli ribosomes, and this methodology have potential to accelerate the understanding of ribosome biogenesis, function, and the development of engineered ribosomes with additional functions.

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