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Intrinsically Disordered Protein Analysis pp 387–398Cite as

Mass Spectrometry Tools for Analysis of Intermolecular Interactions

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 896))

Abstract

The small quantities of protein required for mass spectrometry (MS) make it a powerful tool to detect binding (protein–protein, protein–small molecule, etc.) of proteins that are difficult to express in large quantities, as is the case for many intrinsically disordered proteins. Chemical cross-linking, proteolysis, and MS analysis, combined, are a powerful tool for the identification of binding domains. Here, we present a traditional approach to determine protein–protein interaction binding sites using heavy water (18O) as a label. This technique is relatively inexpensive and can be performed on any mass spectrometer without specialized software.

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Acknowledgement

This work was supported in part by a National Institutes of Health R21 (to J.N.A.) (1R21NS071256), R21 (to C.A.S.) (R21 A1067021), P01 (to C.A.S.) (P01 GM091743), GM 32415 and GM 26788 (to G.A.P. and D.R.), and Fidelity Biosciences Research Initiative (to G.A.P. and D.R.).

Author information

Authors and Affiliations

  1. Department of Biochemistry and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA, USA

    Jared R. Auclair, Dagmar Ringe & Gregory A. Petsko

  2. Department of Chemistry and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA, USA

    Jared R. Auclair & Gregory A. Petsko

  3. Chemistry and Volen Center for Complex Systems, Brandeis University, Waltham, MA, USA

    Jared R. Auclair & Jeffrey N. Agar

  4. Department of Pediatrics, University of Massachusetts Medical School, Worcester, MA, USA

    Mohan Somasundaran

  5. Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA

    Karin M. Green, James E. Evans & Celia A. Schiffer

  6. Department of Proteomics and Mass Spectrometry, University of Massachusetts Medical School, Worcester, MA, USA

    Karin M. Green & James E. Evans

  7. Departments of Chemistry and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA, USA

    Dagmar Ringe

Authors
  1. Jared R. Auclair
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  2. Mohan Somasundaran
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  3. Karin M. Green
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  4. James E. Evans
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  5. Celia A. Schiffer
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  6. Dagmar Ringe
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  7. Gregory A. Petsko
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  8. Jeffrey N. Agar
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Corresponding authors

Correspondence to Jared R. Auclair or Jeffrey N. Agar .

Editor information

Editors and Affiliations

  1. DEPT OF MOLECULAR MED, College of Medicine, University of South Florida, BRUCE B DOWNS BLVD 12901, TAMPA, 33612, Florida, USA

    Vladimir N. Uversky

  2. Dept. Biochemistry & Molecular Biology, Center for Computational Biology &, Indiana University, West 10th Street 410, Indianapolis, 46202, Indiana, USA

    A. Keith Dunker

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Auclair, J.R. et al. (2012). Mass Spectrometry Tools for Analysis of Intermolecular Interactions. In: Uversky, V., Dunker, A. (eds) Intrinsically Disordered Protein Analysis. Methods in Molecular Biology, vol 896. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-3704-8_26

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  • DOI: https://doi.org/10.1007/978-1-4614-3704-8_26

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4614-3703-1

  • Online ISBN: 978-1-4614-3704-8

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