A two-dimensional array for simultaneous sequencing of N- and o-glycans and their glycoforms on specific glycosylation sites

Journal of Proteome Research
Yaron TzurRachel G Lichtenstein

Abstract

Glycans play major roles in living organisms. Thus, essential information is required on diverse glycans, their location, and moieties in proteins, as well as for technology in a high-throughput manner, for improving functional glycomics. In the present study, we describe a new approach involving a 2-D array, which has the potential to fulfill both requirements. The first dimension of the array is composed of various lectins immobilized to a MALDI plate. The second dimension consists of initial proteolysis, then sequential exoglycosidase digestion using highly specific enzymes. The products of such digestions are peptide/glycopeptide mixtures conjugating different glycan fragments from which the exoglycosidase has removed specific terminal residues. Consequently, a series of spectra are obtained when lectin-attached products are analyzed by MALDI-TOF MS. By using well-known glycoproteins and NKp46D2-Ig, a recombinant fusion natural killer receptor with unknown glycans produced in CHO cells, we proved the usefulness of the method, demonstrating rapid and simultaneous determination of N- and O-glycan sequences, their glycan moieties, and subtypes on each of the determined glycosylation sites. This strategy provides a tool that ca...Continue Reading

Citations

May 26, 2011·Molecular & Cellular Proteomics : MCP·Erika LattováBoguslaw Tomanek
Jan 22, 2011·Mass Spectrometry Reviews·Pawel L UrbanRenato Zenobi
Jul 15, 2011·Journal of Pharmaceutical and Biomedical Analysis·Song C KlapoetkeSteven Becht
Sep 17, 2017·Scientific Reports·Marton Szigeti, Andras Guttman

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