Abstract
Technologies to sequence nucleic acids/proteins are widely available, but straightforward methodologies to sequence complex polysaccharides are lacking. We here put forward a strategy to sequence glycosaminoglycans, long linear polysaccharides involved in many biochemical processes. The method is based on the covalent immobilization and (immuno)chemical characterization of only those size-separated saccharides that harbor the original reducing end of the full-length chain. Using this methodology, the saccharide sequence of the chondroitin sulfate chain of the proteoglycan bikunin was determined. The method can be performed in any standard biochemical lab and opens studies to the interaction of complex saccharide sequences with other biomolecules.
Highlights
In addition to nucleic acids and proteins, glycosaminoglycans have been indicated as a class of information-dense biomolecules
We used bioorthogonal reaction pairs (“click chemistry”), labeling the reducing end of glycosaminoglycans with one reaction partner, and transferring them to blot paper functionalized with the other reaction partner
Oligosaccharides were used as model compounds and labeling was evaluated by a shift in migration after polyacrylamide gel electrophoresis (Fig. 2a,b)
Summary
In addition to nucleic acids and proteins, glycosaminoglycans have been indicated as a class of information-dense biomolecules. Fragmentation and in-gel visualization of end-labeled chondroitin sulfate. We functionalized blotting paper with the bioorthogonal reaction partner to allow immobilization of only end-labeled fragments, and expose them in a way allowing (immuno)chemical characterization.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
