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Mass Spectrometric Identification of In Vivo Nitrotyrosine Sites in the Human Pituitary Tumor Proteome

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Book cover Neuroproteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 566))

Summary

The chemically stable tyrosine nitration of a protein involves the addition of a nitro group (–NO2) to the phenolic ring of a tyrosine residue, which may be associated with nervous system physiological and pathological processes. Identification of nitrotyrosine sites on a protein could clarify the functional significance of the modification. Due to the rarity of nitrotyrosine sites in a proteome, tandem mass spectrometry, coupled with different techniques that isolate and enrich nitrotyrosine-containing proteins from a pituitary proteome, is currently the most effective method for site identification. Commercially available nitrotyrosine polyclonal/monoclonal antibodies enable one to detect nitrotyrosine-containing proteins in a two-dimensional gel electrophoresis (2DGE) map, and to preferentially enrich nitrotyrosine-containing proteins with immunoprecipitation. Our present protocols have integrated different isolation/enrichment techniques (2DGE; Western blots; nitrotyrosine immunoaffinity precipitation) and two different tandem mass spectrometry methods (MALDI-MS/MS; ESI-MS/MS) to determine the amino acid sequence of nitrotyrosine-containing peptides that derive from nitrated proteins. Bioinformatics tools are then used to correlate nitrotyrosine sites with a functional domain/motif in order to understand the relationship between tyrosine nitration and the structural/functions of proteins.

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Acknowledgments

The authors acknowledge financial support from NIH (NS 42843 to DMD; RR 16679 to DMD).

Author information

Authors and Affiliations

  1. Charles B. Stout Neuroscience Mass Spectrometry Laboratory, Department of Neurology, University of Tennessee Health Science Center, Memphis, TN, USA

    Xianquan Zhan

  2. Charles B. Stout Neuroscience Mass Spectrometry Laboratory, Departments of Neurology and Molecular Sciences and the Cancer Institute, University of Tennessee Health Science Center, Memphis, TN, USA

    Dominic M. Desiderio

Authors
  1. Xianquan Zhan
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  2. Dominic M. Desiderio
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Correspondence to Xianquan Zhan .

Editor information

Editors and Affiliations

  1. Dept. Anatomy & Neurobiology, Virginia Commonwealth University School of Medicine, Richmond, U.S.A.

    Andrew K. Ottens

  2. Center of Innovative Research, Banyan Biomarkers, Inc. Center of Innovative Research, Alachua, U.S.A.

    Kevin K.W. Wang

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Zhan, X., Desiderio, D.M. (2009). Mass Spectrometric Identification of In Vivo Nitrotyrosine Sites in the Human Pituitary Tumor Proteome. In: Ottens, A., Wang, K. (eds) Neuroproteomics. Methods in Molecular Biology, vol 566. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-562-6_10

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  • DOI: https://doi.org/10.1007/978-1-59745-562-6_10

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-934115-84-8

  • Online ISBN: 978-1-59745-562-6

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