Differential Proteomics Based on Multidimensional Protein Identification Technology to Understand the Biogenesis of Outer Membrane of Escherichia coli.
Cell envelope proteins in bacteria are typically difficult to characterize due to their low abundance, poor solubility, and the problematic isolation of pure surface fraction with only minimal contamination. Here we describe a method for cell membrane fractionation followed by mass spectrometry-based proteomics to analyze and determine protein abundance in bacterial membranes.
Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Milan, Italy.
Alessandra M
AM
Martorana
Istituto di Tecnologie Biomediche, Consiglio Nazionale delle Ricerche, Via Fratelli Cervi 98, Milan, 20090, Italy.
Sara
S
Motta
Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Milan, Italy.
Paola
P
Sperandeo
Istituto di Tecnologie Biomediche, Consiglio Nazionale delle Ricerche, Via Fratelli Cervi 98, Milan, 20090, Italy. pierluigi.mauri@itb.cnr.it.
Pierluigi
P
Mauri
Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Milan, Italy. alessandra.polissi@unimib.it.
Alessandra
A
Polissi
D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002458:Cell Fractionation; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000302:isolation & purification / Q000378:metabolism; D013058:Mass Spectrometry; D040901:Proteomics / Q000379:methods*
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins
Cell fractionation; Gel-free; Gram-negative cell envelope; Membrane proteins; MudPIT; Outer membrane