Analysis of Mucin-Domain Glycoproteins Using Mass Spectrometry.
Curr Protoc
; 4(7): e1100, 2024 Jul.
Article
em En
| MEDLINE
| ID: mdl-38984456
ABSTRACT
Mucin-domain glycoproteins are characterized by their high density of glycosylated serine and threonine residues, which complicates their analysis by mass spectrometry. The dense glycosylation renders the protein backbone inaccessible to workhorse proteases like trypsin, the vast heterogeneity of glycosylation often results in ion suppression from unmodified peptides, and search algorithms struggle to confidently analyze and site-localize O-glycosites. We have made a number of advances to address these challenges, rendering mucinomics possible for the first time. Here, we summarize these contributions and provide a detailed protocol for mass spectrometric analysis of mucin-domain glycoproteins. © 2024 Wiley Periodicals LLC. Basic Protocol 1 Enrichment of mucin-domain glycoproteins Basic Protocol 2 Enzymatic digestion of mucin-domain glycoprotein(s) Basic Protocol 3 Mass spectrometry data collection for O-glycopeptides Basic Protocol 4 Mass spectrometry data analysis of O-glycopeptides.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Espectrometria de Massas
/
Glicoproteínas
/
Mucinas
Limite:
Humans
Idioma:
En
Revista:
Curr Protoc
Ano de publicação:
2024
Tipo de documento:
Article
País de publicação:
EEUU
/
ESTADOS UNIDOS
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ESTADOS UNIDOS DA AMERICA
/
EUA
/
UNITED STATES
/
UNITED STATES OF AMERICA
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US
/
USA