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Enrichment and LC-MS/MS analysis of S-acylated proteins

ABSTRACT: HeLa cells were lysed and free cysteines capped with iodoacetamide. S-acylation was hydrolysed with hydroxylamine and newly revealed Cys allowed to react with an alkyne-tagged probe. Proteins were clicked with azido-biotin, enriched on neutravidin beads, digested with trypsin and desalted.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Cervix Carcinoma

SUBMITTER: EMMANUELLE THINON

LAB HEAD: Emmanuelle Thinon

PROVIDER: PXD062183 | Pride | 2026-06-29

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	IAA-iPrL_Maxquant.txt.zip	Txt
	Legend_PRIDEsubmission.xlsx	Xlsx
	et240328_07_200nl.pepXML	Pepxml
	et240328_07_200nl.raw	Raw
	et240328_08_200nl.pepXML	Pepxml

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Optimization of a method to quantify S-acylation

Project description:HeLa cells were lysed and free cysteines were alkylated with an alkyne-tagged light probe (IAA-BnL or IAA-PrL). S-acylation was hydrolysed with hydroxylamine and newly revealed Cys allowed to react with the heavy probe (IAA-BnL or IAA-PrL). Proteins were clicked with azido-biotin, Adadps-biotin, AazoB or ARB, enriched on neutravidin beads, digested with trypsin and desalted. Different optimisations of the protocol were tested: -8 sets of probes / capture reagent -LC-MS/MS optimisation (gradient, FAIMS, etc) Finally, S-acylation was quantified for a biological replicate of HeLa cells.

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