Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Sci. STKE, 12 June 2001
Vol. 2001, Issue 86, p. pl1
[DOI: 10.1126/stke.2001.86.pl1]

PROTOCOLS

The Biotin Switch Method for the Detection of S-Nitrosylated Proteins

Samie R. Jaffrey, and Solomon H. Snyder

The authors are in the Departments of Neuroscience, Pharmacology and Molecular Sciences, and Psychiatry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. E-mail: jaffrey{at}jhmi.edu; ssnyder {at}jhmi.edu

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.

Citation:
S. R. Jaffrey, S. H. Snyder, The Biotin Switch Method for the Detection of S-Nitrosylated Proteins. Science's STKE (2001), http://stke.sciencemag.org/cgi/content/full/OC_sigtrans;2001/86/pl1.

© 2001 American Association for the Advancement of Science

Citation: S. R. Jaffrey, S. H. Snyder, The Biotin Switch Method for the Detection of S-Nitrosylated Proteins. Sci. STKE 2001, pl1 (2001).

Read the Full Text

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Uterine Smooth Muscle S-Nitrosylproteome in Pregnancy.
C. Ulrich, D. R. Quillici, K. Schegg, R. Woolsey, A. Nordmeier, and I. L. O. Buxton (2012)
Mol. Pharmacol. 81, 143-153
   Abstract »    Full Text »    PDF »
Nitric Oxide and Protein S-Nitrosylation Are Integral to Hydrogen Peroxide-Induced Leaf Cell Death in Rice.
A. Lin, Y. Wang, J. Tang, P. Xue, C. Li, L. Liu, B. Hu, F. Yang, G. J. Loake, and C. Chu (2012)
Plant Physiology 158, 451-464
   Abstract »    Full Text »    PDF »
PNAS Plus: Site-specific and redox-controlled S-nitrosation of thioredoxin.
K. T. Barglow, C. G. Knutson, J. S. Wishnok, S. R. Tannenbaum, and M. A. Marletta (2011)
PNAS 108, E600-E606
   Abstract »    Full Text »    PDF »
SNO-ing at the Nociceptive Synapse?.
I. Tegeder, R. Scheving, I. Wittig, and G. Geisslinger (2011)
Pharmacol. Rev. 63, 366-389
   Abstract »    Full Text »    PDF »
Caveolin-1-eNOS signaling promotes p190RhoGAP-A nitration and endothelial permeability.
M. R. Siddiqui, Y. A. Komarova, S. M. Vogel, X. Gao, M. G. Bonini, J. Rajasingh, Y.-Y. Zhao, V. Brovkovych, and A. B. Malik (2011)
J. Cell Biol. 193, 841-850
   Abstract »    Full Text »    PDF »
Nitric oxide is the primary mediator of cytotoxicity induced by GSH depletion in neuronal cells.
K. Aquilano, S. Baldelli, S. Cardaci, G. Rotilio, and M. R. Ciriolo (2011)
J. Cell Sci. 124, 1043-1054
   Abstract »    Full Text »    PDF »
What can we learn about cardioprotection from the cardiac mitochondrial proteome?.
M. Gucek and E. Murphy (2010)
Cardiovasc Res 88, 211-218
   Abstract »    Full Text »    PDF »
GAPDH regulates cellular heme insertion into inducible nitric oxide synthase.
R. Chakravarti, K. S. Aulak, P. L. Fox, and D. J. Stuehr (2010)
PNAS 107, 18004-18009
   Abstract »    Full Text »    PDF »
Redox Regulatory Mechanism of Transnitrosylation by Thioredoxin.
C. Wu, T. Liu, W. Chen, S.-i. Oka, C. Fu, M. R. Jain, A. M. Parrott, A. T. Baykal, J. Sadoshima, and H. Li (2010)
Mol. Cell. Proteomics 9, 2262-2275
   Abstract »    Full Text »    PDF »
Targeted Quantitation of Site-Specific Cysteine Oxidation in Endogenous Proteins Using a Differential Alkylation and Multiple Reaction Monitoring Mass Spectrometry Approach.
J. M. Held, S. R. Danielson, J. B. Behring, C. Atsriku, D. J. Britton, R. L. Puckett, B. Schilling, J. Campisi, C. C. Benz, and B. W. Gibson (2010)
Mol. Cell. Proteomics 9, 1400-1410
   Abstract »    Full Text »    PDF »
H2S Signals Through Protein S-Sulfhydration.
A. K. Mustafa, M. M. Gadalla, N. Sen, S. Kim, W. Mu, S. K. Gazi, R. K. Barrow, G. Yang, R. Wang, and S. H. Snyder (2009)
Science Signaling 2, ra72
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882