title: Analytic framework for peptidomics applied to large-scale neuropeptide identification.
abstract: Large-scale mass spectrometry-based peptidomics for drug discovery is relatively unexplored because of challenges in peptide degradation and identification following tissue extraction. Here we present a streamlined analytical pipeline for large-scale peptidomics. We developed an optimized sample preparation protocol to achieve fast, reproducible and effective extraction of endogenous peptides from sub-dissected organs such as the brain, while diminishing unspecific protease activity. Each peptidome sample was analysed by high-resolution tandem mass spectrometry and the resulting data set was integrated with publically available databases. We developed and applied an algorithm that reduces the peptide complexity for identification of biologically relevant peptides. The developed pipeline was applied to rat hypothalamus and identifies thousands of neuropeptides and their post-translational modifications, which is combined in a resource format for visualization, qualitative and quantitative analyses.
authors.affiliation: Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Blegdamsvej 3b, DK-2200 Copenhagen, Denmark.|Histology and Imaging, Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Maaloev, Denmark.
authors.forename: Anna
authors.initials: A
authors.lastname: Secher
authors.affiliation: Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Blegdamsvej 3b, DK-2200 Copenhagen, Denmark.
authors.forename: Christian D
authors.initials: CD
authors.lastname: Kelstrup
authors.affiliation: Protein &Peptide Chemistry, Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Maaloev, Denmark.
authors.forename: Kilian W
authors.identifier: http://orcid.org/0000-0001-5303-4448
authors.initials: KW
authors.lastname: Conde-Frieboes
authors.affiliation: Histology and Imaging, Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Maaloev, Denmark.
authors.forename: Charles
authors.initials: C
authors.lastname: Pyke
authors.affiliation: Incretin &Obesity Pharmacology, Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Maaloev, Denmark.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Raun
authors.affiliation: Incretin &Obesity Research, Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Maaloev, Denmark.
authors.forename: Birgitte S
authors.initials: BS
authors.lastname: Wulff
authors.affiliation: Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Blegdamsvej 3b, DK-2200 Copenhagen, Denmark.
authors.forename: Jesper V
authors.identifier: http://orcid.org/0000-0002-4747-4938
authors.initials: JV
authors.lastname: Olsen
mesh_terms: D000465:Algorithms; D000595:Amino Acid Sequence; D000818:Animals; D002853:Chromatography, Liquid; D006358:Hot Temperature; D007031:Hypothalamus / Q000737:chemistry* / Q000378:metabolism; D008297:Male; D008872:Microwaves; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000302:isolation & purification* / Q000378:metabolism; D010477:Perfusion; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry
chemical_list: D009479:Neuropeptides