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The Biotin Switch Method for the Detection of S-Nitrosylated Proteins
Abstract
Many of the effects of nitric oxide are mediated by the direct modification of cysteine residues resulting in an adduct called a nitrosothiol. Here, we describe a novel method for detecting proteins that contain nitrosothiols. In this three-step procedure, nitrosylated cysteines are converted to biotinylated cysteines. Biotinylated proteins can then be detected by immunoblotting or can be purified by avidin-affinity chromatography. We include examples of the detection of S-nitrosylated proteins in brain lysates after in vitro S-nitrosylation, as well as the detection of endogenous S-nitrosothiols in selected neuronal proteins.
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Nitric oxide circulates in mammalian plasma primarily as an S-nitroso adduct of serum albumin.
Stamler, J. S., Jaraki, O., Osborne, J., Simon, D. I., Keaney, J., Vita, J., Singel, D., Valeri, C. R., & Loscalzo, J. (1992) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.89.16.7674 · Cited by 1,238
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Zhang, Y., Fonslow, B. R., Shan, B., Baek, M.-C., & Yates, J. R. (2013) · Chemical Reviews · doi:10.1021/cr3003533 · Cited by 1,491
Thiol chemistry and specificity in redox signaling
Winterbourn, C. C., & Hampton, M. B. (2008) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2008.05.004 · Cited by 1,226
H2S signalling through protein sulfhydration and beyond
Paul, B. D., & Snyder, S. H. (2012) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm3391 · Cited by 853
Self-citationRedox-based regulation of signal transduction: principles, pitfalls, and promises
Janssen-Heininger, Y. M. W., Mossman, B. T., Heintz, N. H., Forman, H. J., Kalyanaraman, B., Finkel, T., Stamler, J. S., Rhee, S. G., & van der Vliet, A. (2008) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2008.03.011 · Cited by 784
Proteomic Identification of S -Nitrosylated Proteins in Arabidopsis
Lindermayr, C., Saalbach, G., & Durner, J. (2005) · Plant Physiology · doi:10.1104/pp.104.058719 · Cited by 734
Metadata
- Authors
- Jaffrey, S. R., & Snyder, S. H.
- Venue
- Science Signaling
- Published
- 2001-06-12
- Volume
- 2001
- Issue
- 86
- Pages
- pl1-pl1
- ISSN
- 1937-9145 (ISSN-L)
- 1945-0877
- OpenAlex
- W4238590243
- Cited by
- 176
- References
- 1
- FWCI
- 0.87
- Type
- journal article
- Language
- en
- Open access
- closed
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
