Resin and Magnetic Nanoparticle-Based Free Radical Probes for Glycan Capture, Isolation, and Structural Characterization
| Autor: | Jesse L. Beauchamp, Daniel A. Thomas, Jinshan Gao, Kimberly Fabijanczuk, Kaylee Gaspar, Jungeun Jasmine Lee, Nikunj Desai |
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| Rok vydání: | 2019 |
| Předmět: |
Glycan
Free Radicals 010402 general chemistry Cleavage (embryo) 01 natural sciences Dithiothreitol Article Analytical Chemistry chemistry.chemical_compound Magnetics Polysaccharides Carbohydrate Conformation Molecule Moiety biology Molecular Structure Chemistry 010401 analytical chemistry Combinatorial chemistry 0104 chemical sciences carbohydrates (lipids) Resins Synthetic Covalent bond Functional group biology.protein Nanoparticles Carbohydrate conformation |
| Zdroj: | Anal Chem |
| ISSN: | 1520-6882 |
| Popis: | By combing the merits of solid supports and free radical activated glycan sequencing (FRAGS) reagents, we develop a multi-functional solid-supported free radical probe (SS-FRAGS), which enables glycan enrichment and characterization. SS-FRAGS comprises a solid support, free radical precursor, disulfide bond, pyridyl, and hydrazine moieties. Thio-activated resin and magnetic nanoparticles (MNPs) are chosen as the solid support to selectively capture free glycans via the hydrazine moiety, allowing for their enrichment and isolation. The disulfide bond acts as a temporary covalent linkage between the solid support and the captured glycan, allowing the release of glycans via the cleavage of the disulfide bond by dithiothreitol. The basic pyridyl functional group provides a site for the formation of a fixed charge, enabling detection by mass spectrometry and avoiding glycan rearrangement during collisional activation. The free radical precursor generates a nascent free radical upon collisional activation and thus simultaneously induces systematic and predictable fragmentation for glycan structure elucidation. A radical-driven glycan deconstruction diagram (R-DECON) is developed to visually summarize the MS(2) results and thus allow for the assembly of the glycan skeleton, making the differentiation of isobaric glycan isomers unambiguous. For application to a real-world sample, we demonstrate the efficacy of the SS-FRAGS by analyzing glycan structures enzymatically cleaved from RNase-B. |
| Databáze: | OpenAIRE |
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