Nat Methods. 2009 Nov;6(11):809-11. Epub 2009 Oct 18.

Enrichment of glycopeptides for glycan structure and attachment site identification.

Nilsson J, Rüetschi U, Halim A, Hesse C, Carlsohn E, Brinkmalm G, Larson G.

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Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, Sweden. jonas.nilsson@clinchem.gu.se

Abstract

We present a method to enrich for glycoproteins from proteomic samples. Sialylated glycoproteins were selectively periodate-oxidized, captured on hydrazide beads, trypsinized and released by acid hydrolysis of sialic acid glycosidic bonds. Mass spectrometric fragment analysis allowed identification of glycan structures, and additional fragmentation of deglycosylated ions yielded peptide sequence information, which allowed glycan attachment site and protein identification. We identified 36 N-linked and 44 O-linked glycosylation sites on glycoproteins from human cerebrospinal fluid.

PMID:: 19838169; [PubMed - indexed for MEDLINE]

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