Improved surfaceome coverage with a label‐free nonaffinity‐purified workflow. Issue 7 (6th March 2017)

• Record Type: Journal Article
• Authors: Glisovic‐Aplenc, Tina; Gill, Saar; Spruce, Lynn A.; Smith, Ian R.; Fazelinia, Hossein; Shestova, Olga; Ding, Hua; Tasian, Sarah K.; Aplenc, Richard; Seeholzer, Steven H.
• Other Names: Aebersold Ruedi guestEditor.; Bensimon Ariel guestEditor.; Collins Ben C. guestEditor.; Ludwig Christina guestEditor.; Sabido Eduard guestEditor.
• Is Part Of: Proteomics. Volume 17:Issue 7(2017)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2017-03-06
• Subjects: GO term -- Label‐free -- Leukemia -- Plasma membrane -- Surfaceome; Proteins -- Separation -- Periodicals; Bioinformatics -- Periodicals; Proteomics -- Periodicals; Genomes -- Periodicals; Molecular genetics -- Periodicals; 572.605
• DOI: 10.1002/pmic.201600344
• Languages: English
• ISSNs: 1615-9853

Abstract

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.