Enrichment of glycopeptides for glycan structure and attachment site identification.
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.
Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, Sweden. jonas.nilsson@clinchem.gu.se
Jonas
J
Nilsson
Ulla
U
Rüetschi
Adnan
A
Halim
Camilla
C
Hesse
Elisabet
E
Carlsohn
Gunnar
G
Brinkmalm
Göran
G
Larson
D002556:Cerebrospinal Fluid Proteins / Q000032:analysis*; D006020:Glycopeptides / Q000032:analysis; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification*; D006031:Glycosylation; D006801:Humans; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry; D040901:Proteomics; D053719:Tandem Mass Spectrometry
D002556:Cerebrospinal Fluid Proteins; D006020:Glycopeptides; D006023:Glycoproteins; D011134:Polysaccharides; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase