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Complementary Proteomic Analysis of Protein Complexes

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 917))

Abstract

Proteomic characterization of protein complexes leverages the versatile platform of liquid chromatography-tandem mass spectrometry to elucidate molecular and cellular signaling processes underlying the dynamic regulation of macromolecular assemblies. Here, we describe a complementary proteomic approach optimized for immunoisolated protein complexes. As the relative complexity, abundance, and physiochemical properties of proteins can vary significantly between samples, we have provided (1) complementary sample preparation workflows, (2) detailed steps for HPLC and mass spectrometric method development, and (3) a bioinformatic workflow that provides confident peptide/protein identification paired with unbiased functional gene ontology analysis. This protocol can also be extended for characterization of larger complexity samples from whole cell or tissue Xenopus proteomes.

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Author information

Authors and Affiliations

  1. Department of Molecular Biology, Princeton University, Princeton, NJ, USA

    Todd M. Greco, Yana Miteva & Ileana M. Cristea

  2. Department of Genetics, University of North Carolina, Chapel Hill, NC, USA

    Frank L. Conlon

Authors
  1. Todd M. Greco
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  2. Yana Miteva
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  3. Frank L. Conlon
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  4. Ileana M. Cristea
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Corresponding author

Correspondence to Ileana M. Cristea .

Editor information

Editors and Affiliations

  1. , Institute of Medical Sciences, University of Aberdeen, FORESTERHILL, ABERDEEN, AB25 2ZD, United Kingdom

    STEFAN HOPPLER

  2. , Department of Biological Science, University of Calgary, University Drive NW 2500, Calgary, T2N 1N4, Canada

    Peter D Vize

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© 2012 Springer Science+Business Media, LLC

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Greco, T.M., Miteva, Y., Conlon, F.L., Cristea, I.M. (2012). Complementary Proteomic Analysis of Protein Complexes. In: HOPPLER, S., Vize, P. (eds) Xenopus Protocols. Methods in Molecular Biology, vol 917. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-992-1_22

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  • DOI: https://doi.org/10.1007/978-1-61779-992-1_22

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-991-4

  • Online ISBN: 978-1-61779-992-1

  • eBook Packages: Springer Protocols

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