Journal
PROTEOMICS
Volume 9, Issue 22, Pages 5046-5055Publisher
WILEY
DOI: 10.1002/pmic.200900033
Keywords
Glycopeptides; Glycoproteomics; Gold nanoparticles; MALDI-QIT-TOF MS; 4-Mercaptophenylboronic acid; On-plate enrichment; Selective enrichment
Funding
- National Basic Research Priorities Program [2007CB91410013]
- National Natural Science Foundation of China [20875017]
- Shanghai Leading Academic Discipline Project [B109]
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In this study, an on-plate-selective enrichment method is developed for fast and efficient glycopeptide investigation. Gold nanoparticles were first spotted and sintered on a stainless-steel plate, then modified with 4-mercaptophenylboronic acid to provide porous substrate with large specific surface and dual functions. These spots were used to selectively capture glycopeptides from peptide mixtures and the captured target peptides could be analyzed by MALDI-MS simply by deposition of 2,5-dihydroxybenzoic acid matrix. Horseradish peroxidase was employed as a standard glycoprotein to investigate the enrichment efficiency. In this way, the enrichment, washing and detection steps can all be fulfilled on a single MALDI target plate. The relatively small sample amount needed, low detection limit and rapid selective enrichment have made this on-plate strategy promising for online enrichment of glycopeptides, which could be applied in high-throughput proteome research.
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