Dataset Information

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

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 a label-free, non-affinity-purified workflow for the enrichment of the plasma membrane proteome and use subsequent bioinformatics tools to select plasma membrane proteins, herein referred to as the surfaceome. Using this methodology, we identified over 2500 cell surface-associated proteins in a human acute myeloid leukemia (AML) cell line. These surface proteins comprised 60% of all detected cellular proteins, a number that substantially exceeds the depth of coverage in previously published studies describing the leukemia surfaceome.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Leukocyte, Cell Culture

DISEASE(S): Acute Leukemia

SUBMITTER: Steven Seeholzer

LAB HEAD: Steven H. Seeholzer

PROVIDER: PXD004801 | Pride | 2017-01-31

REPOSITORIES: pride

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Dataset's files

Source:

			Action	DRS
	U_Human_Reviewed_RefIso_160329.fasta	Fasta
	e140306_RA780_30_01.raw	Raw
	e140306_RA780_30_02.raw	Raw
	e140905_3_RA927_SDC.raw	Raw
	e141121_2_RA927igd3_02.raw	Raw

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Improved surfaceome coverage with a label-free nonaffinity-purified workflow.

Glisovic-Aplenc Tina T Gill Saar S Spruce Lynn A LA Smith Ian R IR Fazelinia Hossein H Shestova Olga O Ding Hua H Tasian Sarah K SK Aplenc Richard R Seeholzer Steven H SH

Proteomics 20170306 7

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 ...[more]

PMID: 28116781

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