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Improved surfaceome coverage with a label-free nonaffinity-purified workflow.
- Source :
-
Proteomics [Proteomics] 2017 Apr; Vol. 17 (7). Date of Electronic Publication: 2017 Mar 06. - Publication Year :
- 2017
-
Abstract
- The proteins of the cellular plasma membrane (PM) perform important functions relating to homeostasis and intercellular communication. Due to its overall low cellular abundance, amphipathic character, and low membrane-to-cytoplasm ratio, the PM proteome has been challenging to isolate and characterize, and is poorly represented in standard LC-MS/MS analyses. In this study, we employ sucrose gradient ultracentrifugation for the enrichment of the PM proteome, without chemical labeling and affinity purification, together with GeLCMS and use subsequent bioinformatics tools to select proteins associated with the PM/cell surface, herein referred to as the surfaceome. Using this methodology, we identify over 1900 cell surface associated proteins in a human acute myeloid leukemia cell line. These surface proteins comprise almost 50% of all detected cellular proteins, a number that substantially exceeds the depth of coverage in previously published studies describing the leukemia surfaceome.<br /> (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Subjects :
- Cell Line, Tumor
Cell Membrane metabolism
Centrifugation, Density Gradient
Chromatography, Liquid
Humans
Leukocytes metabolism
Sucrose chemistry
Tandem Mass Spectrometry
Ultracentrifugation methods
Cell Membrane chemistry
Computational Biology methods
Leukocytes chemistry
Membrane Proteins isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 1615-9861
- Volume :
- 17
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Proteomics
- Publication Type :
- Academic Journal
- Accession number :
- 28116781
- Full Text :
- https://doi.org/10.1002/pmic.201600344