Abstract

A suite of bioactive peptides orchestrates a variety of cellular interactions in the mammalian brain. The bioanalytical strategy known as neuropeptidomics evolved from the quest to globally characterize these important cell–cell signaling peptides. The goal of a neuropeptidomics experiment is to characterize the peptides present in an intact brain, brain region, or even an individual neuron. A neuropeptidomics measurement needs to contend with the large dynamic range and low abundance of the neuropeptides that are present within a background of peptides resulting from the postmortem degradation of high-level ubiquitous proteins. The core components of a successful effort include effective tissue sampling and stabilization, sensitive and robust peptide characterization, and comprehensive data analysis and interpretation. Mass spectrometry (MS) has become the central analytical approach for high-throughput characterization of the brain peptidome because of its capability to detect, identify, and quantify known and unknown peptides with high confidence. Robust fractionation techniques, such as two-dimensional liquid chromatography (LC), are commonly used in conjunction with MS to enhance investigation of the peptidome. Identification and characterization of peptides is more complex when neuropeptide prohormone genes have not been annotated in an unbiased manner. This chapter outlines techniques and describes protocols for three different experimental designs that combine MS with LC, each aimed at high-throughput discovery of peptides in brain tissue. Further, we describe the currently available bioinformatics tools for automatic query of the experimental data against existing protein databases, manual retrieval of structural information from raw MS data, and label-free quantitation.

Original languageEnglish (US)
Title of host publicationNeuromethods
PublisherHumana Press Inc.
Pages161-177
Number of pages17
DOIs
StatePublished - Jan 1 2019

Publication series

NameNeuromethods
Volume146
ISSN (Print)0893-2336
ISSN (Electronic)1940-6045

Fingerprint

Brain
Peptides
Mass spectrometry
Mass Spectrometry
Liquid chromatography
Neuropeptides
Liquid Chromatography
Throughput
Tissue
Protein Databases
Information Storage and Retrieval
Bioinformatics
Fractionation
Computational Biology
Design of experiments
Neurons
Labels
Proteins
Research Design
Stabilization

Keywords

  • Bioinformatics
  • Hormone
  • Liquid chromatography
  • Mass spectrometry
  • Neuropeptide
  • Neuropeptidome

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Psychiatry and Mental health

Cite this

Xie, F., Anapindi, K. D. B., Romanova, E. V., & Sweedler, J. V. (2019). Neuropeptidomics of the mammalian brain. In Neuromethods (pp. 161-177). (Neuromethods; Vol. 146). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9662-9_14

Neuropeptidomics of the mammalian brain. / Xie, Fang; Anapindi, Krishna D.B.; Romanova, Elena Vikentievna; Sweedler, Jonathan V.

Neuromethods. Humana Press Inc., 2019. p. 161-177 (Neuromethods; Vol. 146).

Research output: Chapter in Book/Report/Conference proceedingChapter

Xie, F, Anapindi, KDB, Romanova, EV & Sweedler, JV 2019, Neuropeptidomics of the mammalian brain. in Neuromethods. Neuromethods, vol. 146, Humana Press Inc., pp. 161-177. https://doi.org/10.1007/978-1-4939-9662-9_14
Xie F, Anapindi KDB, Romanova EV, Sweedler JV. Neuropeptidomics of the mammalian brain. In Neuromethods. Humana Press Inc. 2019. p. 161-177. (Neuromethods). https://doi.org/10.1007/978-1-4939-9662-9_14
Xie, Fang ; Anapindi, Krishna D.B. ; Romanova, Elena Vikentievna ; Sweedler, Jonathan V. / Neuropeptidomics of the mammalian brain. Neuromethods. Humana Press Inc., 2019. pp. 161-177 (Neuromethods).
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