Isolation and characterization of glycosylated neuropeptides.
Animals
Chromatography, Liquid
/ instrumentation
Glycopeptides
/ chemistry
Glycosylation
Humans
Isomerism
Mass Spectrometry
/ instrumentation
Models, Molecular
Neuropeptides
/ chemistry
Polysaccharides
/ analysis
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
/ instrumentation
Tandem Mass Spectrometry
/ instrumentation
Characterization
Derivatization
ESI
EThcD
Enrichment
Glycopeptide
Glycosylation
LC-MS/MS
MALDI
Separation
Journal
Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271
Informations de publication
Date de publication:
2019
2019
Historique:
entrez:
14
10
2019
pubmed:
14
10
2019
medline:
3
6
2020
Statut:
ppublish
Résumé
Glycosylation is one of the most ubiquitous and complex post-translational modifications (PTMs). It plays pivotal roles in various biological processes. Studies at the glycopeptide level are typically considered as a downstream work resulting from enzymatic digested glycoproteins. Less attention has been focused on glycosylated endogenous signaling peptides due to their low abundance, structural heterogeneity and the lack of enabling analytical tools. Here, protocols are presented to isolate and characterize glycosylated neuropeptides utilizing nanoflow liquid chromatography coupled with mass spectrometry (LC-MS). We first demonstrate how to extract neuropeptides from raw tissues and perform further separation/cleanup before MS analysis. Then we describe hybrid MS methods for glycosylated neuropeptide profiling and site-specific analysis. We also include recommendations for data analysis to identify glycosylated neuropeptides in crustaceans where a complete neuropeptide database is still lacking. Other strategies and future directions are discussed to provide readers with alternative approaches and further unravel biological complexity rendered by glycosylation.
Identifiants
pubmed: 31606074
pii: S0076-6879(19)30312-X
doi: 10.1016/bs.mie.2019.07.032
pmc: PMC7170398
mid: NIHMS1557623
pii:
doi:
Substances chimiques
Glycopeptides
0
Neuropeptides
0
Polysaccharides
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
147-202Subventions
Organisme : NIDDK NIH HHS
ID : R01 DK071801
Pays : United States
Organisme : NIA NIH HHS
ID : RF1 AG052324
Pays : United States
Organisme : NCI NIH HHS
ID : U01 CA231081
Pays : United States
Informations de copyright
© 2019 Elsevier Inc. All rights reserved.
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