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Determining Copper and Zinc Content in Superoxide Dismutase Using Electron Capture Dissociation Under Native Spray Conditions

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Proteoform Identification

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2500))

Abstract

Localizing metal binding to specific sites in proteins remains a challenging analytical problem in vitro and in vivo. Although metal binding can be maintained by “native” electrospray ionization with intact proteins for quantitation by mass spectrometry, subsequent fragmentation of proteins with slow-heating methods like collision-induced dissociation (CID) can scramble and detach metals. In contrast, electron capture dissociation (ECD) fragmentation produces highly localized bond cleavage that is well known to preserve posttranslational modifications. We show how a newly available ECD tool that can be retrofitted on standard QTOF mass spectrometers allows the sites of copper and zinc binding to be localized in the antioxidant enzyme Cu, Zn superoxide dismutase (SOD1). The loss of zinc from Cu, Zn SOD1 has been shown to induce motor neuron death and could have a causal role in the fatal neurodegenerative disease, amyotrophic lateral sclerosis (ALS). The methods described enable copper loss to be distinguished from zinc using distinct ECD fragments of SOD1 and are broadly applicable to other metalloproteins.

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Author information

Authors and Affiliations

  1. Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA

    Rachel Franklin & Joseph S. Beckman

  2. e-MSion Inc., Corvallis, OR, USA

    Michael Hare & Joseph S. Beckman

Authors
  1. Rachel Franklin
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  2. Michael Hare
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  3. Joseph S. Beckman
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Corresponding author

Correspondence to Joseph S. Beckman .

Editor information

Editors and Affiliations

  1. Department of Chemistry, Michigan State University, East Lansing, MI, USA

    Liangliang Sun

  2. Deming Department of Medicine, School of Medicine, Tulane University, New Orleans, LA, USA

    Xiaowen Liu

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Franklin, R., Hare, M., Beckman, J.S. (2022). Determining Copper and Zinc Content in Superoxide Dismutase Using Electron Capture Dissociation Under Native Spray Conditions. In: Sun, L., Liu, X. (eds) Proteoform Identification. Methods in Molecular Biology, vol 2500. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2325-1_14

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  • DOI: https://doi.org/10.1007/978-1-0716-2325-1_14

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2324-4

  • Online ISBN: 978-1-0716-2325-1

  • eBook Packages: Springer Protocols

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