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Tandem mass spectra of glycan substructures enable the multistage mass spectrometric identification of determinants on oligosaccharides
- Source :
- Rapid Communications in Mass Spectrometry. 27:931-939
- Publication Year :
- 2013
- Publisher :
- Wiley, 2013.
-
Abstract
- RATIONALE: Glycosylation of proteins and lipids affects many biological processes, such as host–pathogen interactions, cell communication, and initiation of the immune responses. Terminal glycan substructures, or determinants, often govern the function or recognition of the carrier glycoconjugate and modulate these processes. In this study we describe a strategy using multistage mass spectrometry to identify and confirm these glycan substructures. METHODS: An online tandem mass spectrometry (MS2) spectral fragment library of glycan substructures that typically occur at the non-reducing terminus of glycoconjugates was created to enable the easier identification and confirmation of glycan determinants on oligosaccharides released from glycoproteins. Oligosaccharides were separated by porous graphitized carbon capillary chromatography and analysed by ion trap MS. Candidate product ions that constitute the glycan substructure mass were identified in the MS2 product ion spectrum, and used as the precursor ion for subsequent MS3 fragmentation. The resulting MS3 spectrum was matched against the MS2 spectral fragment library to identify the glycan substructure(s) that comprise the parent oligosaccharide. RESULTS: Thirty biologically important terminal glycan determinants commonly observed on glycoconjugates were fragmented by positive and negative ion mass spectrometry and the MS2 product ion masses manually annotated and stored in the UniCarb-DB online database. Negative ion tandem mass spectra were especially useful in assigning isobaric glycan structures. We have applied this strategy for the identification of the sulphation, blood group antigens and sialic acid linkages on complex N-and O-glycans released from glycoproteins. CONCLUSIONS: We show the potential of these glycan substructure MS2 spectra in the negative ionization mode to facilitate the assignment of determinants on N- and O-linked glycans released from glycoproteins. Comparing the structural feature ions of known glycan reference substructures assists in the annotation of complex glycan product ion spectra, and can remove the need for other orthogonal confirmation analyses such as sequential glycosidase digestion. Copyright © 2013 John Wiley & Sons, Ltd.
- Subjects :
- chemistry.chemical_classification
Glycan
Glycosylation
biology
Glycoconjugate
Organic Chemistry
Oligosaccharide
Mass spectrometry
Tandem mass spectrometry
Analytical Chemistry
Sialic acid
carbohydrates (lipids)
chemistry.chemical_compound
Biochemistry
chemistry
biology.protein
Ion trap
Spectroscopy
Subjects
Details
- ISSN :
- 09514198
- Volume :
- 27
- Database :
- OpenAIRE
- Journal :
- Rapid Communications in Mass Spectrometry
- Accession number :
- edsair.doi...........e1f7b6f1249b76fad283f6e4e9abf9b3