One-dimensional capillary liquid chromatographic separation coupled with tandem mass spectrometry unveils the escherichia coli proteome on a microarray scale

Mio Iwasaki, Shohei Miwa, Tohru Ikegami, Masaru Tomita, Nobuo Tanaka, Yasushi Ishihama

研究成果: Article

108 引用 (Scopus)

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We successfully identified the proteome expressed in Escherichia coli (E. coli) cells on a microarray scale using one-dimensional capillary liquid chromatography-tandem mass spectrometry (LC-MS/MS) with a 350 cm long, 100 μm i. d., monolithic silica-C18 capillary column. E. coli tryptic digest (4 μg) was injected onto the column, and a 41 h gradient was applied with a flow rate of 500 nL/min at less than 20 MPa. In total, 22 196 nonredundant tryptic peptides from 2602 proteins, including 830 membrane proteins, were identified from the E. coli cells (triplicate analysis), in which an equivalent number of genes was detected by transcriptome analysis. Approximately a 5-fold larger peak response on average was obtained in this system, compared with that obtained by conventional capillary LC-MS/MS analysis with a 15 cm long, 3 μm diameter C18 silica particle-packed column. The higher response suggests that the influence of ionization suppression was drastically reduced by the high-efficiency separation on the long monolithic silica column coupled with the shallow gradient. Because this high-resolution system does not require any additional separation prior to LC-MS/MS, this "one-shot" proteomics approach can simplify the workflow of shotgun proteomics and minimize the sample amount, as well as reduce the total analysis time, despite the use of prolonged shallow gradient elution.

元の言語English
ページ(範囲)2616-2620
ページ数5
ジャーナルAnalytical chemistry
82
発行部数7
DOI
出版物ステータスPublished - 2010 4 1

ASJC Scopus subject areas

  • Analytical Chemistry

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