Isolation and characterization of glycosylated neuropeptides
- Resource Type
- Authors
- Yang Liu; Qinjingwen Cao; Lingjun Li
- Source
- Methods in enzymology. 626
- Subject
- Models, Molecular
Glycosylation
030303 biophysics
Neuropeptide
Computational biology
Mass Spectrometry
Article
03 medical and health sciences
chemistry.chemical_compound
Isomerism
Polysaccharides
Tandem Mass Spectrometry
Lc ms ms
Animals
Humans
chemistry.chemical_classification
0303 health sciences
Neuropeptides
Ms analysis
Glycopeptides
Structural heterogeneity
Glycopeptide level
carbohydrates (lipids)
chemistry
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Glycoprotein
Chromatography, Liquid
- Language
- ISSN
- 1557-7988
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 neuropeptides 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.