Analytic framework for peptidomics applied to large-scale neuropeptide identification

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 pre...

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Vydáno v:	Nature communications Ročník 7; číslo 1; s. 11436 - 10
Hlavní autoři:	Secher, Anna, Kelstrup, Christian D., Conde-Frieboes, Kilian W., Pyke, Charles, Raun, Kirsten, Wulff, Birgitte S., Olsen, Jesper V.
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	London Nature Publishing Group UK 04.05.2016 Nature Publishing Group Nature Portfolio
Témata:	631/378/340 631/45/611 631/61 64 82 82/16 82/58 82/81 Algorithms Amino Acid Sequence Animals Brain research Chromatography Chromatography, Liquid Datasets Hot Temperature Humanities and Social Sciences Hypothalamus Hypothalamus - chemistry Hypothalamus - metabolism Male Mass spectrometry Microwaves Molecular Sequence Annotation Molecular Sequence Data multidisciplinary Neuropeptides Neuropeptides - isolation & purification Neuropeptides - metabolism Obesity Peptides Perfusion Phosphorylation Protein Processing, Post-Translational Proteins Proteomics - methods Protocol R&D Rats Rats, Sprague-Dawley Research & development Sample preparation Science Science (multidisciplinary) Scientific imaging Software Tandem Mass Spectrometry Denmark
ISSN:	2041-1723, 2041-1723
On-line přístup:	Získat plný text
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Shrnutí:	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. Neuropeptide research is challenged by technical difficulties in identifying new bioactive peptides. Here the authors present an analytical pipeline for large-scale peptidomics applied to the rat hypothalamus, identifying thousands of endogenous neuropeptides and their post-translational modifications.
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 These authors contributed equally to this work.
ISSN:	2041-1723 2041-1723
DOI:	10.1038/ncomms11436