An optimized protocol for isolation of S-nitrosylated proteins from C. elegans

Post-translational modification by S-nitrosylation regulates numerous cellular functions and impacts most proteins across phylogeny. We describe a protocol for isolating S-nitrosylated proteins (SNO-proteins) from C. elegans, suitable for assessing SNO levels of individual proteins and of the global...

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Bibliographic Details
Published in:STAR protocols 2021-06, Vol.2 (2), p.100547-100547, Article 100547
Main Authors: Seth, Puneet, Premont, Richard T., Stamler, Jonathan S.
Format: Article
Language:English
Subjects:
Animals
Caenorhabditis elegans - chemistry
Caenorhabditis elegans Proteins - analysis
Caenorhabditis elegans Proteins - chemistry
Caenorhabditis elegans Proteins - isolation & purification
Model Organisms
Nitrosation
Protein Biochemistry
Proteome - analysis
Proteome - chemistry
Proteome - isolation & purification
Proteomics
Proteomics - methods
Protocol
S-Nitrosothiols
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Summary:Post-translational modification by S-nitrosylation regulates numerous cellular functions and impacts most proteins across phylogeny. We describe a protocol for isolating S-nitrosylated proteins (SNO-proteins) from C. elegans, suitable for assessing SNO levels of individual proteins and of the global proteome. This protocol features efficient nematode lysis and SNO capture, while protection of SNO proteins from degradation is the major challenge. This protocol can be adapted to mammalian tissues. For complete information on the generation and use of this protocol, please refer to Seth et al. (2019). [Display omitted] •Protocol for isolating S-nitrosylated proteins from C. elegans•Assesses global changes in S-nitroso-protein levels•Also assesses S-nitrosylation changes in individual proteins of interest•Protocol is adaptable to mammalian tissues Post-translational modification by S-nitrosylation regulates numerous cellular functions and impacts most proteins across phylogeny. We describe a protocol for isolating S-nitrosylated proteins (SNO-proteins) from C. elegans, suitable for assessing SNO levels of individual proteins and of the global proteome. This protocol features efficient nematode lysis and SNO capture, while protection of SNO proteins from degradation is the major challenge. This protocol can be adapted to mammalian tissues.
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2021.100547