Dataset Information

Proteome-wide detection of cysteine nitrosylation targets and motifs using bio-orthogonal cleavable linker based enrichment and switch technique (Cys-BOOST)

ABSTRACT: Cysteine (Cys) reversible post-translational modifications (PTMs) are emerging as important players in cellular signaling and redox homeostasis. Here we present Cys-BOOST a novel strategy for LC-MS/MS based quantitative analysis of reversibly modified Cys using switch technique, enrichment via bio-orthogonal cleavable linker and quantification using tandem mas tag (TMT) reagents. We performed direct comparison of Cys-BOOST (n=3) with iodo-TMT (n=3) by analyzing the total CysCys from HeLa cell extracts. As a result higher sensitivity (25,019 vs 9,966 Cys peptides), specificity (98 vs 74 %) and technical reproducibility were obtained by Cys-BOOST. In addition, the application of Cys-BOOST for the analysis of Cys nitrosylation (SNO) in S-nitrosoglutathione (GSNO) treated and non-treated HeLa cell extracts lead to the identification of unprecedented number of SNO proteins (3,537), SNO peptides (9,314) and unique SNO sites (8,304). Based on the quantitative data we describe SNO consensus motifs for endogenous SNO and SNO sites with differential reactivity to GSNO. Collectively, our findings suggest Cys-BOOST as a concurrent method of choice for Cys PTM analysis.

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Ruzanna Mnatsakanyan

LAB HEAD: René P. Zahedi

PROVIDER: PXD011131 | Pride | 2019-05-21

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	HeLa-GSNO_Cys-BOOST.mzML	Mzml
	HeLa-GSNO_Cys-BOOST.mzid.gz	Mzid
	HeLa-GSNO_Cys-BOOST.pdResult	Other
	HeLa-GSNO_Cys-BOOST.pride.mgf.gz	Mgf
	HeLa-GSNO_FT-1ug.mzML	Mzml

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Proteome-wide detection of S-nitrosylation targets and motifs using bioorthogonal cleavable-linker-based enrichment and switch technique.

Mnatsakanyan Ruzanna R Markoutsa Stavroula S Walbrunn Kim K Roos Andreas A Verhelst Steven H L SHL Zahedi René P RP

Nature communications 20190516 1

Cysteine modifications emerge as important players in cellular signaling and homeostasis. Here, we present a chemical proteomics strategy for quantitative analysis of reversibly modified Cysteines using bioorthogonal cleavable-linker and switch technique (Cys-BOOST). Compared to iodoTMT for total Cysteine analysis, Cys-BOOST shows a threefold higher sensitivity and considerably higher specificity and precision. Analyzing S-nitrosylation (SNO) in S-nitrosoglutathione (GSNO)-treated and non-treate ...[more]

PMID: 31097712

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