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Site-Specific Proteomic Mapping of Modified Cysteine Residues

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Functional Disulphide Bonds

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1967))

Abstract

The wide reactivity of the thiol group enables the formation of a number of chemically and biologically distinct posttranslational modifications. Proteins within nearly all major families undergo some form of cysteine modification and the modifications are associated with regulatory functions across many biological processes. However, the susceptibility of thiols to redox shifts, as well as the labile nature of most thiol modifications, renders detection difficult. Analysis difficulties are compounded further in complex protein mixtures due to the typical low abundance of cysteine modifications under normal physiological conditions. Here we describe methods for the analysis of three cysteine modifications: nitrosylation, glutathionylation, and S-acylation. The three methods use the same organic mercury-conjugated agarose resin as an enrichment platform. To date, over 2154 sites on 1446 proteins have been identified between the three modifications using this method. Using equivalent processing, enrichment, and analytical methods has enabled a more comprehensive picture of the redox proteome landscape.

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Author information

Authors and Affiliations

  1. Department of Pediatrics, Children’s Hospital of Philadelphia Research Institute, Philadelphia, PA, USA

    Neal S. Gould

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  1. Neal S. Gould
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Corresponding author

Correspondence to Neal S. Gould .

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Editors and Affiliations

  1. The Centenary Institute, NHMRC Clinical Trials Centre, Sydney Medical School, University of Sydney, Sydney, NSW, Australia

    Philip Hogg

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Gould, N.S. (2019). Site-Specific Proteomic Mapping of Modified Cysteine Residues. In: Hogg, P. (eds) Functional Disulphide Bonds. Methods in Molecular Biology, vol 1967. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9187-7_11

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  • DOI: https://doi.org/10.1007/978-1-4939-9187-7_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9186-0

  • Online ISBN: 978-1-4939-9187-7

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