A Two-Dimensional Array for Simultaneous Sequencing of N- And O-Glycans and Their Glycoforms on Specific Glycosylation Sites

Yaron Tzur, Amir Markovich, Rachel G. Lichtenstein

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

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-lg, 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 can rapidly explore glycan structures and might contribute to a better understanding of process- and disease-related glycoproteins.

Original languageEnglish
Pages (from-to)1188-1198
Number of pages11
JournalJournal of Proteome Research
Volume7
Issue number3
DOIs
StatePublished - 1 Mar 2008

Keywords

  • MALDI-TOF MS
  • N-glycan
  • NKp46D2-lg
  • O-glycan
  • Two-dimensional array

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry (all)

Fingerprint

Dive into the research topics of 'A Two-Dimensional Array for Simultaneous Sequencing of N- And O-Glycans and Their Glycoforms on Specific Glycosylation Sites'. Together they form a unique fingerprint.

Cite this