title: Enrichment of plasma membrane proteins using nanoparticle pellicles: comparison between silica and higher density nanoparticles.
abstract: Proteomic and other characterization of plasma membrane proteins is made difficult by their low abundance, hydrophobicity, frequent carboxylation, and dynamic population. We and others have proposed that underrepresentation in LC-MS/MS analysis can be partially compensated by enriching the plasma membrane and its proteins using cationic nanoparticle pellicles. The nanoparticles increase the density of plasma membrane sheets and thus enhance separation by centrifugation from other lysed cellular components. Herein, we test the hypothesis that the use of nanoparticles with increased densities can provide enhanced enrichment of plasma membrane proteins for proteomic analysis. Multiple myeloma cells were grown and coated in suspension with three different pellicles of three different densities and both pellicle coated and uncoated suspensions analyzed by high-throughput LC-MS/MS. Enrichment was evaluated by the total number and the spectral counts of identified plasma membrane proteins.
authors.affiliation: Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, United States.
authors.forename: Waeowalee
authors.initials: W
authors.lastname: Choksawangkarn
authors.forename: Sung-Kyoung
authors.initials: SK
authors.lastname: Kim
authors.forename: Joe R
authors.initials: JR
authors.lastname: Cannon
authors.forename: Nathan J
authors.initials: NJ
authors.lastname: Edwards
authors.forename: Sang Bok
authors.initials: SB
authors.lastname: Lee
authors.forename: Catherine
authors.initials: C
authors.lastname: Fenselau
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D002498:Centrifugation; D002853:Chromatography, Liquid; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D009101:Multiple Myeloma / Q000378:metabolism / Q000473:pathology; D053758:Nanoparticles; D012822:Silicon Dioxide; D053719:Tandem Mass Spectrometry
chemical_list: D008565:Membrane Proteins; D012822:Silicon Dioxide