Lipid rafts, microdomain heterogeneity and inter-organelle contacts: impacts on membrane preparation for proteomic studies.
In recent years, there has been considerable interest in mapping the protein content of isolated organelles using mass spectrometry. However, many subcellular compartments are highly dynamic with diverse and intricate architectures that are not always preserved during membrane isolation procedures. Furthermore, lateral heterogeneities in intra-membrane lipid and protein concentrations underlie the formation of membrane microdomains, trafficking vesicles and inter-membrane contacts. These complexities in membrane organisation have important consequences for the design of membrane preparation strategies and test the very concept of organelle purity. We illustrate how some of these biological considerations are relevant to membrane preparation and assess the numerous potential pitfalls in attempting to purify organelles from mammalian cells.
Centre for Molecular Cell Biology, UCL, Institute of Liver and Digestive Health, Royal Free Campus, UCL, London NW3 2PF, United Kingdom.
Shane
S
Minogue
Mark G
MG
Waugh
D016477:Artifacts; D002458:Cell Fractionation / Q000379:methods* / Q000592:standards; D006801:Humans; D007425:Intracellular Membranes / Q000737:chemistry*; D013058:Mass Spectrometry; D008563:Membrane Lipids / Q000737:chemistry; D021962:Membrane Microdomains / Q000737:chemistry*; D008565:Membrane Proteins / Q000032:analysis*; D015388:Organelles / Q000737:chemistry*; D021381:Protein Transport; D040901:Proteomics; D013347:Subcellular Fractions / Q000737:chemistry*; D022161:Transport Vesicles / Q000737:chemistry
D008563:Membrane Lipids; D008565:Membrane Proteins