A Bacterial Mannose Binding Lectin as a Tool for the Enrichment of C- and O-Mannosylated Peptides
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Mass spectrometry (MS) easily detects C-mannosylated peptides from purified proteins but not from complex biological samples. Enrichment of specific glycopeptides by lectin affinity prior to MS analysis has been widely applied to support glycopeptide identification but was until now not available for C-mannosylated peptides. Here, we used the α-mannose-specific Burkholderia cenocepacia lectin A (BC2L-A) and show that, in addition to its previously demonstrated high-mannose N-glycan binding capability, this lectin is able to retain C- and O-mannosylated peptides. Besides testing binding abilities to standard peptides, we applied BC2L-A affinity to enrich C-mannosylated peptides from complex samples of tryptic digests of HEK293 and MCF10A whole cell extracts, which led to the identification of novel C-mannosylation sites. In conclusion, BC2L-A enabled specific enrichment of C- and O-mannosylated peptides and might have superior properties over other mannose binding lectins for this purpose.
Originalsprog | Engelsk |
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Tidsskrift | Analytical Chemistry |
Vol/bind | 94 |
Udgave nummer | 20 |
Sider (fra-til) | 7329–7338 |
ISSN | 0003-2700 |
DOI | |
Status | Udgivet - 2022 |
Bibliografisk note
Funding Information:
This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) (FOR 2509 Projektnummer: 289991887), Project-ID BA4091/6-2 (to H.B.), BU2920/2-2 (to F.F.R.B.), RA2992/1-2 (to E.R.), and STR 443/5-1 and STR 443/6-1 (to S.S.). A.I. was supported by PIA Glyco@Alps (ANR-15-IDEX-02) and Labex Arcane/CBH-EUR-GS (ANR-17-EURE-0003). L.P., S.Y.V., and A.H. were supported by the Danish National Research Foundation (DNRF107) and a research grant (00025438) from VILLUM FONDEN.
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© 2022 American Chemical Society
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