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Characterization of the Phosphoproteome and Sialoproteome in Brain Tissues by Mass Spectrometry

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Current Proteomic Approaches Applied to Brain Function

Part of the book series: Neuromethods ((NM,volume 127))

Abstract

Mass spectrometry is an essential tool for the characterization of proteins within neuroscience. The development of faster instruments enables neuroscientists to investigate a large proportion of the proteome in the brain in only short analysis time. Yet, a detailed functional investigation of the intrinsic biochemical processes of the brain by evaluation of the post-translational modifications in proteins is still missing. Phosphorylation and N-linked glycosylation are important protein modifications within the brain as they are involved in neural development, neurotransmission, neurite extension, and synaptic plasticity. Although the importance of these protein modifications is undoubtable for the brain functionality, only a few global protein modification datasets have been generated so far in the neuroscience field. This is due to the higher difficulties to sensitively and specifically enrich these low abundant protein modifications from the high abundant nonmodified peptides and from the very lipid-rich brain material.

Here, we describe how a highly selective, sensitive, low hands-on-time and cost-effective simultaneous enrichment of phosphorylated peptides, sialylated N-linked glycopeptides as well as intact sialylated N-linked glycopeptides and unmodified peptides from the same biological sample can be applied to bridge this gap in neuroscience, exemplified by a proteomic characterization of the murine brain growth spurt.

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Abbreviations

AAA:

Amino acid analysis

ACN:

Acetonitrile

AMPA:

Alpha-amino-2-hydroxy-5-methyl-4-isoxazole propionic acid receptor

CDG:

Congenital disorder of glycosylation

CNS:

Central nervous system

DTT:

Dithiothreitol

HCD:

Higher collisional-induced dissociation

HILIC:

Hydrophilic interaction liquid chromatography

IAA:

Iodoacetamide

IMAC:

Iron metal affinity chromatography

LTD:

Long-term depression

LTP:

Long-term potentiation

MALDI:

Matrix-assisted laser desorption ionization

MeOH:

Methanol

mGluR:

Metabotropic G-protein-coupled glutamate receptor

MS:

Mass spectrometry

nLC-MS/MS:

Nano liquid chromatography-Tandem mass spectrometry

NMDAR:

N-Methyl-d-aspartate receptor

PTM:

Post-translational modification

RT:

Room temperature

SIMAC:

Sequential elution from IMAC

SPE:

Solid phase extraction

TEAB:

Triethylammonium bicarbonate

TFA:

Trifluoroacetic acid

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Author notes

  1. María Ibáñez-Vea and Stefan J. Kempf contributed equally to this work.

Authors and Affiliations

  1. Immunomodulation Group, Navarrabiomed Biomedical Research Center, Navarra Institute for Health Research (IdiSNA), Pamplona, Spain

    María Ibáñez-Vea

  2. Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230, Odense, Denmark

    Stefan J. Kempf & Martin R. Larsen

Authors
  1. María Ibáñez-Vea
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  2. Stefan J. Kempf
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  3. Martin R. Larsen
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Corresponding author

Correspondence to Martin R. Larsen .

Editor information

Editors and Affiliations

  1. Clinical Neuroproteomics Unit, Navarrabiomed, Navarra Health Department, Public University of Navarra, Proteored-Institute of Health Carlos III (ISCIII), Navarra Institute for Health Research (IdiSNA), Pamplona, Spain

    Enrique Santamaría

  2. Clinical Neuroproteomics Unit, Navarrabiomed, Navarra Health Department, Public University of Navarra, Proteored-Institute of Health Carlos III (ISCIII), Navarra Institute for Health Research (IdiSNA), Pamplona, Spain

    Joaquín Fernández-Irigoyen

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Ibáñez-Vea, M., Kempf, S.J., Larsen, M.R. (2017). Characterization of the Phosphoproteome and Sialoproteome in Brain Tissues by Mass Spectrometry. In: Santamaría, E., Fernández-Irigoyen, J. (eds) Current Proteomic Approaches Applied to Brain Function. Neuromethods, vol 127. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7119-0_12

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  • DOI: https://doi.org/10.1007/978-1-4939-7119-0_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7118-3

  • Online ISBN: 978-1-4939-7119-0

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