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

STAR Protoc. 2021 May 19;2(2):100547. doi: 10.1016/j.xpro.2021.100547. eCollection 2021 Jun 18.

Abstract

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).

Keywords: Model Organisms; Protein Biochemistry; Proteomics.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Caenorhabditis elegans / chemistry*
  • Caenorhabditis elegans Proteins* / analysis
  • Caenorhabditis elegans Proteins* / chemistry
  • Caenorhabditis elegans Proteins* / isolation & purification
  • Nitrosation
  • Proteome* / analysis
  • Proteome* / chemistry
  • Proteome* / isolation & purification
  • Proteomics / methods*
  • S-Nitrosothiols

Substances

  • Caenorhabditis elegans Proteins
  • Proteome
  • S-Nitrosothiols