Peptidomics pp 191-206 | Cite as

Identification and Relative Quantification of Neuropeptides from the Endocrine Tissues

  • Kurt Boonen
  • Steven J. Husson
  • Bart Landuyt
  • Geert Baggerman
  • Eisuke Hayakawa
  • Walter H.M.L. Luyten
  • Liliane Schoofs
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 615)

Abstract

Endocrine tissues like the pituitary, hypothalamus and islets of Langerhans are rich in bioactive peptides. These are used for intercellular signalling and are involved in regulation of almost all physiological processes. Peptidomics is the comprehensive analysis of peptides in tissues, fluids and cells. Peptidomics applied to (neuro-)endocrine tissues aims therefore to identify as many bioactive peptides as possible. Peptidomics of (neuro-)endocrine tissues requires an integrated approach that consists of careful sample handling, peptide separation techniques, mass spectrometry and bioinformatics. Here we describe the methods for isolation and dissection of endocrine tissues, the extraction of bioactive peptides and further sample handling and identification of peptides by mass spectrometry and hyphenated techniques. We also present a straightforward method for the comparison of relative levels of bioactive peptides in these endocrine tissues under varying physiological conditions. The latter helps to elucidate functions of the bioactive peptides.

Key words

Neuropeptide Mus musculus peptidomics pituitary hypothalamus islets of Langerhans endocrine pancreas MALDI-TOF/TOF MS Q-TOF MS two-dimensional HPLC nanoLC 

Notes

Acknowledgments

K. Boonen and B. Landuyt are supported by grants of Institute for the Promotion of Innovation through Science and Technology (I.W.T.)-Flanders. S. J. Husson is a postdoctoral fellow of the Research Foundation Flanders (F.W.O.-Vlaanderen). The authors also acknowledge the Interfacultary Centre for Proteomics and Metabolomics “Prometa”, K.U. Leuven.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Kurt Boonen
    • 1
  • Steven J. Husson
    • 2
  • Bart Landuyt
    • 1
  • Geert Baggerman
    • 3
  • Eisuke Hayakawa
    • 1
  • Walter H.M.L. Luyten
    • 4
  • Liliane Schoofs
    • 1
  1. 1.Functional Genomics and Proteomics Research Unit, Department of BiologyK.U. LeuvenLeuvenBelgium
  2. 2.Functional Genomics and Proteomics, Department of BiologyK.U. LeuvenLeuvenBelgium
  3. 3.ProMeta, Interfacultary Center for Proteomics and Metabolomics, K.U. LeuvenDiepenbeekBelgium
  4. 4.Department Woman and Child, Faculty of MedicineK.U.LeuvenLeuvenBelgium

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