期刊
BIOLOGY-BASEL
卷 11, 期 4, 页码 -出版社
MDPI
DOI: 10.3390/biology11040506
关键词
glycosaminoglycans; mass spectrometry; chondroitin sulfate
类别
资金
- German Research Foundation through the Collaborative Research Centre 1340
- Sonnenfeld foundation
- Deutsche Gesellschaft fur Kardiologie (Otto-Hess-Promotionsstipendium)
In this study, an optimized strategy was presented to analyze glycosaminoglycan disaccharides via MALDI-TOF mass spectrometry, successfully identifying sulfation positions and patterns through fragmentation experiments.
Simple Summary Glycosaminoglycans (GAGs) are considered to be the most difficult type of glycoconjugate to analyze as they are constituted of linear long polysaccharidic chains having molecular weights reaching up to several million daltons. Structural analysis of glycosaminoglycans from biological samples is a long and work-extensive procedure due to the many preparation steps involved. In addition, so far, only few research articles have been dedicated to the analysis of GAGs by means of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) because their ionization can be problematic due to the presence of labile sulfate groups. In this work, we present an optimized strategy to analyze GAG disaccharides via MALDI-TOF mass spectrometry, using a fast workflow that does not require purification after enzymatic cleavage. For the first time, we show that it was possible to identify the sulfation position in disaccharides obtained from GAG via fragmentation experiments. This proof of concept is illustrated via the analysis of chondroitin/dermatan sulfate disaccharides of atherosclerotic lesions, in which we were able to identify their monosulfation patterns. Glycosaminoglycans (GAGs) are considered to be the most difficult type of glycoconjugates to analyze as they are constituted of linear long polysaccharidic chains having molecular weights reaching up to several million daltons. Bottom-up analysis of glycosaminoglycans from biological samples is a long and work-extensive procedure due to the many preparation steps involved. In addition, so far, only few research articles have been dedicated to the analysis of GAGs by means of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) because their intact ionization can be problematic due to the presence of labile sulfate groups. In this work, we had the aim of exploring the sulfation pattern of monosulfated chondroitin/dermatan sulfate (CS/DS) disaccharides in human tissue samples because they represent the most abundant form of sulfation in disaccharides. We present here an optimized strategy to analyze on-target derivatized CS/DS disaccharides via MALDI-TOF-MS using a fast workflow that does not require any purification after enzymatic cleavage. For the first time, we show that MALDI-TOF/TOF experiments allow for discrimination between monosulfated CS disaccharide isomers via specific fragments corresponding to glycosidic linkages and to cross-ring cleavages. This proof of concept is illustrated via the analysis of CS/DS disaccharides of atherosclerotic lesions of different histological origins, in which we were able to identify their monosulfation patterns.
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