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Membrane Proteomics pp 295–310Cite as

Liquid Chromatography MALDI MS/MS for Membrane Proteome Analysis

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 528))

Abstract

Liquid chromatography (LC) can be combined with matrix-assisted laser-desorption ionization (MALDI) mass-spectrometry (MS) by using automated off-line fraction collection of eluates onto a MALDI plate. Commercial tandem MS systems are available for generating product ion spectra of MALDI-produced peptide ions. The MALDI MS/MS spectra can be searched against a proteome database for protein identification. In this chapter, a protocol of sequential solubilization and digestion of membrane proteins involving methanol- and SDS-assisted trypsin digestion and microwave-assisted acid hydrolysis is presented. The process of LC eluate deposition onto a MALDI plate along with practical considerations for achieving optimal performance of eluate deposition is described. Issues related to MALDI MS and MS/MS spectral acquisition are discussed. Database searching and manual inspection of MS/MS spectra of singly charged MALDI-produced peptide ions for positive protein identification are also addressed.

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Author information

Authors and Affiliations

  1. Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada

    Nan Wang, J. Bryce Young & Liang Li

Authors
  1. Nan Wang
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  2. J. Bryce Young
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  3. Liang Li
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Editor information

Editors and Affiliations

  1. Kennedy Institute of Rheumatology Division, Faculty of Medicine Imperial College London, UK

    Matthew J. Peirce  & Robin Wait  & 

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

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Wang, N., Young, J.B., Li, L. (2009). Liquid Chromatography MALDI MS/MS for Membrane Proteome Analysis. In: Peirce, M.J., Wait, R. (eds) Membrane Proteomics. Methods in Molecular Biology™, vol 528. Humana Press. https://doi.org/10.1007/978-1-60327-310-7_21

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  • DOI: https://doi.org/10.1007/978-1-60327-310-7_21

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-309-1

  • Online ISBN: 978-1-60327-310-7

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