Improved plasma membrane proteome coverage with a label-free non-affinity-purified workflow.

TitleImproved plasma membrane proteome coverage with a label-free non-affinity-purified workflow.
Publication TypeJournal Article
Year of Publication2017
AuthorsGlisovic-Aplenc T, Gill S, Spruce LA, Smith IR, Fazelinia H, Shestova O, Ding H, Tasian SK, Aplenc R, Seeholzer SH
JournalProteomics
Date Published2017 Jan 23
ISSN1615-9861
Abstract

The proteins of the cellular plasma membrane 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 plasma membrane proteome has been challenging to isolate and characterize, and is poorly represented in standard liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses. In this study, we employ sucrose gradient ultracentrifugation for the enrichment of the plasma membrane proteome, without chemical labeling and affinity purification, together with GeLCMS and use subsequent bioinformatics tools to select plasma membrane proteins, herein referred to as the surfaceome. Using this methodology, we identify over 1900 cell surface-associated proteins in a human acute myeloid leukemia (AML) 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. This article is protected by copyright. All rights reserved.

DOI10.1002/pmic.201600344
Alternate JournalProteomics
PubMed ID28116781