Dataset Information

Fatty-acylation in RAW264.7 - Bioorthogonal Chemical Reporters for Monitoring Unsaturated Fatty-Acylated Proteins

ABSTRACT: Dietary unsaturated fatty acids beneficially affect human health, in part by modulating the immune system, but the mechanism is not completely understood. Given that unsaturated fatty acids have been shown to be covalently incorporated into a small subset of proteins, we designed three alkyne-tagged chemical reporters of unsaturated fatty acids, alk-16:1, alk-17:1 and alk-18:1, to explore the generality and diversity of this protein modification. Following cell lysis, proteins labelled with the reporters could be captured by azido-functionalized reagents via CuAAC for fluorescence detection or enrichment for proteomics analysis. These reporters label many proteins in mammalian cells and can be incorporated site-specifically, notably on Cys residues. Quantitative proteomics analysis (n= 4 biological replicates) of LPS/IFN-gamma stimulated RAW264.7 labelled with oleic acid (control), alk-16 (palmitic acid chemical reporter), alk-16:1, alk-17:1 and alk-18:1, revealed that unsaturated fatty acids modify similar protein targets to saturated fatty acids, including several immune proteins. Interestingly, some proteins can be differentially labeled with unsaturated and saturated fatty acid.

INSTRUMENT(S): LTQ Orbitrap XL

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Macrophage Cell Line, Macrophage

DISEASE(S): Leukemia

SUBMITTER: Emmanuelle Thinon

LAB HEAD: Howard C. Hang

PROVIDER: PXD003545 | Pride | 2018-10-26

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	MS151605XL_Emmanuelle_Hang_16-A.raw	Raw
	MS151605XL_Emmanuelle_Hang_16-B.raw	Raw
	MS151605XL_Emmanuelle_Hang_16-C.raw	Raw
	MS151605XL_Emmanuelle_Hang_16-D.raw	Raw
	MS151605XL_Emmanuelle_Hang_17-A.raw	Raw

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Bioorthogonal Chemical Reporters for Monitoring Unsaturated Fatty-Acylated Proteins.

Thinon Emmanuelle E Percher Avital A Hang Howard C HC

Chembiochem : a European journal of chemical biology 20160729 19

Dietary unsaturated fatty acids, such as oleic acid, have been shown to be covalently incorporated into a small subset of proteins, but the generality and diversity of this protein modification has not been studied. We synthesized unsaturated fatty-acid chemical reporters and determined their protein targets in mammalian cells. The reporters can induce the formation of lipid droplets and be incorporated site-specifically onto known fatty-acylated proteins and label many proteins in mammalian cel ...[more]

PMID: 27350074

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