The membrane complexome of a new Pseudomonas strain during growth on lysogeny broth medium and medium containing glucose or phenol

• Enrichment of the membrane proteome from Pseudomonas sp. strain phDV1 by two washing steps with 5 mM EDTA and 0.1% lauryl sarcosinate.
• Solubilization using n-dodecyl-β-maltoside (DM) and separation of the native membrane protein complexes by BN PAGE.
• Combination of the BN-PAGE with Tricine-SDS-PAGE and protein identification of tryptic peptides by mass spectrometry.

In this study, we have performed a systematic analysis of Pseudomonas sp. strain phDV1 membrane protein complexes by growing the strain in lysogeny broth medium, and medium containing glucose or phenol as sole carbon sources. In order to study the membrane complexome, we developed an approach for the extraction and the analysis of the membrane protein complexes in native conditions. Our strategy involves (a) enrichment of the membrane proteome from Pseudomonas sp. strain phDV1 by two washing steps; (b) solubilization using n-dodecyl-β-maltoside; (c) a combination of BN-PAGE with Tricine-SDS-PAGE; and (d) protein identification of tryptic peptides by mass spectrometry.

Figure optionsDownload as PowerPoint slide