Autor: |
Chongsaritsinsuk J; Department of Chemistry, Yale University, New Haven, CT., Rangel-Angarita V; Department of Chemistry, Yale University, New Haven, CT., Mahoney KE; Department of Chemistry, Yale University, New Haven, CT., Lucas TM; Department of Chemistry, Yale University, New Haven, CT., Enny OM; Department of Chemistry, Yale University, New Haven, CT., Katemauswa M; Department of Chemistry, Yale University, New Haven, CT., Malaker SA; Department of Chemistry, Yale University, New Haven, CT. |
Jazyk: |
angličtina |
Zdroj: |
BioRxiv : the preprint server for biology [bioRxiv] 2024 Jul 07. Date of Electronic Publication: 2024 Jul 07. |
DOI: |
10.1101/2024.07.06.602348 |
Abstrakt: |
Protein glycosylation is a complex post-translational modification that is generally classified as N- or O-linked. Site-specific analysis of glycopeptides is accomplished with a variety of fragmentation methods, depending on the type of glycosylation being investigated and the instrumentation available. For instance, collisional dissociation methods are frequently used for N-glycoproteomic analysis with the assumption that one N-sequon exists per tryptic peptide. Alternatively, electron-based methods are indispensable for O-glycosite localization. However, the presence of simultaneously N- and O-glycosylated peptides could suggest the necessity of electron-based fragmentation methods for N-glycoproteomics, which is not commonly performed. Thus, we quantified the prevalence of N- and O-glycopeptides in mucins and other glycoproteins. A much higher frequency of co-occupancy within mucins was detected whereas only a negligible occurrence occurred within non-mucin glycoproteins. This was demonstrated from analyses of recombinant and/or purified proteins, as well as more complex samples. Where co-occupancy occurred, O-glycosites were frequently localized to the Ser/Thr within the N-sequon. Additionally, we found that O-glycans in close proximity to the occupied Asn were predominantly unelaborated core 1 structures, while those further away were more extended. Overall, we demonstrate electron-based methods are required for robust site-specific analysis of mucins, wherein co-occupancy is more prevalent. Conversely, collisional methods are generally sufficient for analyses of other types of glycoproteins. |
Databáze: |
MEDLINE |
Externí odkaz: |
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