Dataset Information

FluoroTRAQ: Quantitative Analysis of Protein S-nitrosylation through Fluorous Solid-phase Extraction Combining with iTRAQ by Mass Spectrometry

ABSTRACT: S-nitrosylation is an important post-translational modification which occurs on cysteine in organisms. S-nitrosylation regu-lates signal transduction in many cell processes and aberrant S-nitrosylated (SNO-) proteins are closely related to cardiovas-cular and neurological diseases. However, quantitative analysis of the S-nitrosylation at the proteome level remains challeng-ing due to the low abundance of the modification. Herein, we developed a novel approach called FluoroTRAQ, which com-bined the fluorous solid-phase extraction of SNO-peptides and the iTRAQ labeling for the quantitative analysis of SNO-proteome with high sensitivity and specificity. The new analytical strategy is successfully applied to monitor of the dynamic changes of SNO-proteome in cells. A quantitative dataset of SNO-proteome in human umbilical vein endothelial cell (HUVEC) after in vivo S-nitrosoglutathione (GSNO) stimulation was revealed. Among them, several new SNO-proteins in-cluding DHX9, GOT2 et al. are reported and validated by biotin switch assay.

ORGANISM(S): Homo Sapiens

SUBMITTER: Haojie Lu

PROVIDER: PXD020657 | iProX | Thu Jul 30 00:00:00 BST 2020

REPOSITORIES: iProX

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Publications

FluoroTRAQ: Quantitative Analysis of Protein S-Nitrosylation through Fluorous Solid-Phase Extraction Combining with iTRAQ by Mass Spectrometry.

Zhang Cheng C Xu Yaoyao Y Wang Guoli G Fang Caiyun C Bao Huimin H Zhang Ying Y Lu Haojie H

Analytical chemistry 20201111 23

S-Nitrosylation is an important post-translational modification that occurs on cysteine amino acid and regulates signal transduction in diverse cell processes. Dysregulation of protein nitrosylation has shown close association with cardiovascular and neurological diseases, thus demanding further precise and in-depth understanding. Mass spectrometry-based proteomics has been the method of choice for analyzing S-nitrosylated (SNO-) proteins. However, due to their extremely low expression level and ...[more]

PMID: 33174720

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Project description:Hypermethylation of helicase-like transcription factor (HLTF) in colorectal cancer (CRC) cells occurs more frequently in men than women. Progressive epigenetic silencing of HLTF in tumor cells is accompanied by negligible expression in the tumor microenvironment (TME). Cell line-derived xenografts were established in control (Hltf+/+) and Hltf-deleted male Rag2-/-IL2rg-/- mice by direct orthotopic microinjection of HLTF+/+HCT116 cells into the submucosa of the cecum. Combinatorial induction of IL6 and S100A8/A9 in the Hltf-deleted TME with ICAM-1 and IL8 in the primary tumor activated a positive feedback loop. The proinflammatory niche produced a major shift in CDX metastasis to peritoneal dissemination compared to controls. Inducible nitric oxide (iNOS) gene expression and transactivation of the iNOS-S100A8/A9 signaling complex in Hltf-deleted TME reprogrammed the human S-nitroso-proteome. S-nitroso (SNO) proteins POTEE, TRIM52 and UN45B are S-nitrosylated on the conserved I/L-X-C-X2-D/E motif indicative of iNOS-S100A8/A9-mediated S-nitrosylation. 2D-DIGE and protein identification by MALDI-TOF/TOF mass spectrometry authenticated S-nitrosylation of 53 individual cysteines in half-site motifs (I/L-X-C or C-X-X-D/E) in CDX tumors. POTEE-SNO in CDX tumors is both a general S-nitrosylation target and an iNOS-S100A8/A9 site-specific (Cys638) target in the Hltf-deleted TME. REL is an example of convergence of transcriptomic-S-nitroso-proteomic signaling. The gene is transcriptionally activated in CDX tumors with an Hltf-deleted TME, and REL-SNO (Cys143) is found in primary CDX tumors and all metastatic sites. Primary CDX tumors from Hltf-deleted TME shared 60% of their S-nitroso-proteome with all metastatic sites. Forty percent of SNO-proteins from primary CDX tumors were variably expressed at metastatic sites. Global S-nitrosylation of proteins in pathways related to cytoskeleton and motility is strongly implicated in the metastatic dissemination of CDX tumors. Hltf-deletion from the TME plays a major role in the pathogenesis of inflammation and links protein S-nitrosylation in primary CDX tumors with spatiotemporal continuity in metastatic progression even when the tumor cells express HLTF.

Dataset Information

FluoroTRAQ: Quantitative Analysis of Protein S-nitrosylation through Fluorous Solid-phase Extraction Combining with iTRAQ by Mass Spectrometry

Publications

FluoroTRAQ: Quantitative Analysis of Protein S-Nitrosylation through Fluorous Solid-Phase Extraction Combining with iTRAQ by Mass Spectrometry.

Similar Datasets

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