Dataset Information

Proteomic analysis of S-nitrosylated nuclear proteins in rat cortical neurons

ABSTRACT: S-nitrosothiol Resin Assisted Capture(SNORAC) coupled with mass spectrometry analysis has been used to study S-nitrosylated proteins in rat cortical neurons. Proteins were captured on the resin by covalently attaching their former S-NO moiety. Then, nistrosylated proteins were eluted by on-bead digestion. Formerly S-nitrosylated peptides were also eluted from the beads in a second step, and analyzed separately for assessment of the nitrosylation site (qualitative analysis). Out of over 600 nitrosyltated proteins, 131 proteins have never been shown to be S-nitrosylated in any system and 612 are new targets of S-nitrosylation in cortical neurons. The site(s) of Snitrosylation were also identified for 59% of the targets.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Brain, Neuron Of Cerebral Cortex

SUBMITTER: Marco Gaspari

LAB HEAD: Marco Gaspari

PROVIDER: PXD009071 | Pride | 2018-07-11

REPOSITORIES: Pride

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Publications

Proteomic analysis of S-nitrosylated nuclear proteins in rat cortical neurons.

Smith Jacob G JG Aldous Sarah G SG Andreassi Catia C Cuda Giovanni G Gaspari Marco M Riccio Antonella A

Science signaling 20180703 537

Neurons modulate gene expression in response to extrinsic signals to enable brain development, cognition, and learning and to process stimuli that regulate systemic physiological functions. This signal-to-gene communication is facilitated by posttranslational modifications such as S-nitrosylation, the covalent attachment of a nitric oxide (NO) moiety to cysteine thiols. In the cerebral cortex, S-nitrosylation of histone deacetylase 2 (HDAC2) is required for gene transcription during neuronal dev ...[more]

PMID: 29970601

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Project description:Human prion diseases are fatal neurodegenerative disorders characterized by neuronal damage in brain. Protein S-nitrosylation, the covalent adduction of a NO to cysteine, plays a role in human brain biology, and brain dysfunction is a prominent feature of pPrion disease, yet the direct brain targets of S-nitrosylation are largely unknown. We described the first proteomic analysis of global S-nitrosylation in brain tissues of sporadic Creutzfeldt–Jakob disease (sCJD), fatal familial insomnia (FFI) and genetic CJD with a substitution of valine for glycine at codon 114 of the prion protein gene (G114V gCJD) accompanying with normal control with isobaric tags for relative and absolute quantitation (iTRAQ) combined with a nano-HPLC/Q Exactive Mass spectrometry platform. In parallel, we used several approaches to provide quality control for the experimentally defined S-nitrosylated proteins. Total 1509 S-nitrosylated proteins (SNO-proteins) were identified, and cerebellum tissues appeared to contain more commonly differentially expressed SNO-proteins (DESPs) than cortex of sCJD, FFI and gCJD. Three selected SNO-proteins were verified by Western blots, consistent with proteomics assays. Gene ontology analysis showed that more up-regulated DESPs were involved in metabolism, cell cytoskeleton/structure and immune system both in cortex and cerebellum, while more down-regulated ones in both regions were involved in cell cytoskeleton/structure, cell-cell communication and miscelaneous function protein. Pathway analysis suggested that systemic lupus erythematosus, pathogenic Escherichia coli infection, extracellular matrix-receptor interaction were the most commonly affected pathways, which were identified from at least two different diseases. Using STRING database, the network of immune system and cell cytoskeleton and structure were commonly identified in the context of the up-regulated and down-regulated DESPs, respectively, both in cortex and cerebellum. Our study thus have implications for understanding the molecular mechanisms of human prion diseases related to abnormal protein S-nitrosylation and pave the way for future studies focused on potential biomarkers for the diagnosis and therapy of human prion diseases.

Dataset Information

Proteomic analysis of S-nitrosylated nuclear proteins in rat cortical neurons

Publications

Proteomic analysis of S-nitrosylated nuclear proteins in rat cortical neurons.

Similar Datasets

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