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An optimized protocol for isolation of S-nitrosylated proteins from C. elegans
Puneet Seth1, Richard T Premont1,2, Jonathan S Stamler1,2
1Institute for Transformative Molecular Medicine and Department of Medicine, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
STAR Protocols
|June 7, 2021
Summary
This study presents a new protocol for isolating S-nitrosylated proteins (SNO-proteins) from C. elegans. The method efficiently captures SNO-proteins and can be adapted for mammalian tissues.
Area of Science:
- Biochemistry
- Molecular Biology
- Proteomics
Background:
- Post-translational modifications, such as S-nitrosylation, are crucial for regulating cellular functions.
- S-nitrosylation impacts a wide range of proteins across different species.
- Understanding S-nitrosylation is key to deciphering complex cellular signaling pathways.
Purpose of the Study:
- To describe a novel protocol for the isolation of S-nitrosylated proteins (SNO-proteins) from the nematode C. elegans.
- To provide a method for assessing both individual and global SNO-protein levels.
- To offer a protocol adaptable for use with mammalian tissues.
Main Methods:
- Development of an efficient lysis and SNO-capture technique for C. elegans.
- Implementation of strategies to protect SNO-proteins from degradation during isolation.
- Adaptation of the protocol for application to mammalian tissue samples.
Main Results:
- Successful isolation of S-nitrosylated proteins from C. elegans.
- The protocol allows for the assessment of SNO-protein levels at both individual and proteome-wide scales.
- Demonstrated adaptability of the protocol for mammalian tissues.
Conclusions:
- The described protocol offers an effective means to study S-nitrosylation in C. elegans.
- This method facilitates the investigation of S-nitrosylation's role in cellular processes.
- The protocol's adaptability broadens its utility for research in various biological systems.
