Systematic Analysis of Native Membrane Protein Complexes in Escherichia coli

• Published in: Journal of proteome research 2011-04, Vol.10 (4), p.1848-1859
• Main Authors: Maddalo, Gianluca, Stenberg-Bruzell, Filippa, Götzke, Hansjörg, Toddo, Stephen, Björkholm, Patrik, Eriksson, Hanna, Chovanec, Peter, Genevaux, Pierre, Lehtiö, Janne, Ilag, Leopold L, Daley, Daniel O
• Format: Article
• Language: English
• Subjects: Biochemistry; biokemi; BN-PAGE; cell envelope; Chromatography, Ion Exchange - methods; Electrophoresis, Gel, Two-Dimensional - methods; Escherichia coli; Escherichia coli - chemistry; Escherichia coli Proteins - analysis; Escherichia coli Proteins - classification; Escherichia coli Proteins - isolation & purification; Mass Spectrometry - methods; Medicin och hälsovetenskap; membrane protein; Membrane Proteins - analysis; Membrane Proteins - classification; Membrane Proteins - isolation & purification; Molecular Chaperones - analysis; Molecular Chaperones - classification; Molecular Chaperones - isolation & purification; Molecular Weight; Multiprotein Complexes - chemistry; Multiprotein Complexes - isolation & purification; Phylogeny; PpiD; protein complex; proteome; Proteome - analysis; Proteomics - methods; YfgM
• ISSN: 1535-3893; 1535-3907; 1535-3907
• DOI: 10.1021/pr101105c

The cell envelope of Escherichia coli is an essential structure that modulates exchanges between the cell and the extra-cellular milieu. Previous proteomic analyses have suggested that it contains a significant number of proteins with no annotated function. To gain insight into these proteins and the general organization of the cell envelope proteome, we have carried out a systematic analysis of native membrane protein complexes. We have identified 30 membrane protein complexes (6 of which are novel) and present reference maps that can be used for cell envelope profiling. In one instance, we identified a protein with no annotated function (YfgM) in a complex with a well-characterized periplasmic chaperone (PpiD). Using the guilt by association principle, we suggest that YfgM is also part of the periplasmic chaperone network. The approach we present circumvents the need for engineering of tags and protein overexpression. It is applicable for the analysis of membrane protein complexes in any organism and will be particularly useful for less-characterized organisms where conventional strategies that require protein engineering (i.e., 2-hybrid based approaches and TAP-tagging) are not feasible.