Proteomics of total membranes and subcellular membranes

Abstract

Membrane proteins are key molecules in the cell and are important targets for drug development. Much effort has, therefore, been directed towards research of this group of proteins, but their hydrophobic nature can make working with them challenging. Here we discuss methodologies used in the study of the membrane proteome, specifically discussing approaches that circumvent technical issues specific to the membrane. In addition, we review several techniques used for visualization, qualification, quantitation and localization of membrane proteins. The combination of the techniques we describe holds great promise to allow full characterization of the membrane proteome and to map the dynamic changes within it essential for cellular function.

Comparative Skeletal Muscle Proteomics Using Two-Dimensional Gel Electrophoresis

“…These abnormal electrophoretic mobility patterns of particular proteins cause a certain degree of 2D streaking, which can be minimized by (i) decreasing the total amount of protein loading; (ii) using very large gel systems with a higher discriminatory capacity and/or (iii) applying optimized pre-fractionation techniques to decisively decrease sample complexity [95,96,97,98]. Artifacts can be kept to a minimum using 50 to 200 μg of total protein in first dimension gels.…”

Mass spectrometry-based proteomics and peptidomics for systems biology and biomarker discovery

“…Numerous efforts have been made to improve the solubility and enrichment of membrane proteins during sample preparation. Several comprehensive studies recently covered the commonly used technologies in membrane proteomics and different strategies that circumvent technical issues specific to the membrane (Gilmore and Washburn, 2010; Griffin and Schnitzer, 2011; Groen and Lilley, 2010; Helbig et al, 2010; Weekes et al, 2010). Recently, Sun et al .…”

Mass spectrometry-based proteomics and peptidomics for systems biology and biomarker discovery

“…Numerous efforts have been made to improve the solubility and enrichment of membrane proteins during sample preparation. Several comprehensive studies recently covered the commonly used technologies in membrane proteomics and different strategies that circumvent technical issues specific to the membrane (Gilmore and Washburn, 2010;Griffin and Schnitzer, 2011;Groen and Lilley, 2010;Helbig et al, 2010;Weekes et al, 2010). Recently, Sun et al reported using 1-butyl-3-methyl imidazolium tetrafluoroborate (BMIM BF4), an ionic liquid (IL), as a sample preparation buffer for the analysis of integral membrane proteins (IMPs) by microcolumn reversed phase liquid chromatography (3RPLC)-electrospray ionization tandem mass spectrometry (ESI-MS/MS).…”