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Current Proteomic Approaches Applied to Brain Function pp 29–42Cite as

Application of Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) to Monitor Olfactory Proteomes During Alzheimer’s Disease Progression

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

Abstract

Olfactory impairment is a common early feature in several neurodegenerative diseases, including Alzheimer’s disease (AD). One of the main brain regions involved in the processing of olfactory information is the olfactory bulb (OB). In this chapter, we describe the use of isobaric tags for relative and absolute quantification (iTRAQ) to study the OB proteome during the neurodegenerative process in AD subjects. These chemical tags label all peptides in a protein digest via free amines at the peptide N-terminus and on the side chain of lysine residues. Labeled samples are then pooled and analyzed simultaneously using mass spectrometry (MS). Since these tags are isobaric, the intensity of each peak is the sum of the intensity of the peptide from all samples, thus enhancing sensitivity in MS. Similarly, upon peptide fragmentation, amino acid sequence ions also show this summed intensity. However, the distinct distribution of isotopes in the tags is such that when the tags fragment, a tag-specific reporter ion is released. The relative amount of peptide in each of the labeled samples will be represented by the relative intensities of these ions. In summary, this chapter describes the experimental procedure followed to analyzed human OB samples from AD subjects with the aim to increase the understanding of the molecular mechanisms that underlie neurodegeneration in this brain region.

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Acknowledgments

This work was funded by grants from the Spanish Ministry of Economy and Competitiveness (MINECO) (Ref. SAF2014-59340-R), Department of Economic Development from Government of Navarra (Ref. PC025), and Obra Social la Caixa to E.S. A.G.M. is supported by PEJ-2014-A-61949 (MINECO). The proteomics unit of Navarrabiomed is a member of Proteored, PRB2-ISCIII, and is supported by grant PT13/0001 of the PE I+D+I 2013–2016 funded by ISCIII and FEDER.

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Author notes

  1. Andrea González Morales and Mercedes Lachén-Montes contributed equally to this work.

Authors 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

    Andrea González Morales, Mercedes Lachén-Montes, Enrique Santamaría & Joaquín Fernández-Irigoyen

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

    María Ibáñez-Vea

Authors
  1. Andrea González Morales
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  2. Mercedes Lachén-Montes
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  3. María Ibáñez-Vea
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  4. Enrique Santamaría
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  5. Joaquín Fernández-Irigoyen
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Corresponding author

Correspondence to Joaquín Fernández-Irigoyen .

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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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Morales, A.G., Lachén-Montes, M., Ibáñez-Vea, M., Santamaría, E., Fernández-Irigoyen, J. (2017). Application of Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) to Monitor Olfactory Proteomes During Alzheimer’s Disease Progression. 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_3

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  • DOI: https://doi.org/10.1007/978-1-4939-7119-0_3

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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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