Dataset Information

Cell surface mapping and monitoring of cell surface dynamics

ABSTRACT: Here, we present a simple and robust N-linked glycoproteome enrichment protocol optimized for deep coverage of the plasma membrane proteome. The procedure was applied to characterize the cell surface proteome of 15 standard laboratory human cell lines and three primary cell types. We observed significant differences in receptor diversity and transporter abundances between primary cells and corresponding cell lines. Relative quantification using isobaric mass tagging enabled quantitative monitoring of dynamic processes on the cell surface in multiplexed experiments. We for the first time report a comprehensive analysis of the profound remodeling of the plasma membrane proteome during monocyte to macrophage differentiation of THP-1 cells. Treatment of these cells with the kinase inhibitor dasatinib (Sprycel) during differentiation severely compromised macrophage differentiation due to an off-target activity resulting in substantial morphological and functional changes and the loss of many macrophage-associated proteins.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): B Cell, T Cell, Monocyte, Permanent Cell Line Cell, Cell Culture, Macrophage

SUBMITTER: Maria Faelth Savitski

LAB HEAD: Christian Eberl

PROVIDER: PXD005846 | Pride | 2017-03-27

REPOSITORIES: Pride

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	00864_F1_R1_P0102545B02_TMT10.raw.rar	Raw
	00864_F1_R1_P0102545B03_TMT10.raw.rar	Raw
	00864_F1_R1_P0102545B04_TMT10.raw.rar	Raw
	00864_F1_R1_P0102545B05_TMT10.raw.rar	Raw
	00864_F1_R1_P0102545B06_TMT10.raw.rar	Raw

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Publications

Monitoring Cell-surface <i>N</i>-Glycoproteome Dynamics by Quantitative Proteomics Reveals Mechanistic Insights into Macrophage Differentiation.

Kalxdorf Mathias M Gade Stephan S Eberl H Christian HC Bantscheff Marcus M

Molecular & cellular proteomics : MCP 20170323 5

The plasma membrane proteome plays a crucial role in inter- and intracellular signaling, cell survival, and cell identity. As such, it is a prominent target for pharmacological intervention. The relatively low abundance of this subproteome in conjunction with challenging extractability and solubility still hampers its comprehensive analysis. Here, we combined a chemical glycoprotein-tagging strategy with mass spectrometry to enable comprehensive analysis of the cell-surface glycoproteome. To ben ...[more]

PMID: 28336715

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