Quantitative analysis of O-GlcNAcylation in combination with isobaric tag labeling and chemoenzymatic enrichment.
Bioorg Med Chem Lett
; 27(22): 5022-5026, 2017 11 15.
Article
em En
| MEDLINE
| ID: mdl-29029932
ABSTRACT
Protein O-GlcNAcylation regulates various biological processes, and is associated with several diseases. Therefore, the development of quantitative proteomics is important for understanding the mechanisms of O-GlcNAc-related diseases. We previously reported selective enrichment of O-GlcNAcylated peptides, which provided high-selectivity and effective release by a novel thiol-alkyne and thiol-disulfide exchange. Here, we describe a new approach using initial isobaric tag labeling for relative quantification followed by enrichment and ß-elimination/Michael addition with dithiothreitol for identification of both proteins and modification sites. The approach was validated using model proteins and peptides. This novel strategy could be used for quantitative O-GlcNAcome of biological samples.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Peptídeos
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Acetilglucosamina
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Proteômica
Limite:
Animals
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article