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Screening oligosaccharide libraries against lectins using the proxy protein electrospray ionization mass spectrometry assay

  • Author(s) / Creator(s)
  • An electrospray ionization mass spectrometry (ESI-MS) assay for screening carbohydrate libraries against lectins is described. The assay is based on the proxy protein ESI-MS method, which combines direct ESI-MS protein–ligand binding measurements and competitive protein binding, to simultaneously detect and quantify protein–carbohydrate interactions. Specific interactions between components of the library and the target protein (PT) are identified from changes in the relative abundances (as measured by ESI-MS) of the carbohydrate complexes of a proxy protein (Pproxy), which binds to all components of the library with known affinity, upon addition of PT to the solution. The magnitude of the change in relative abundance of a given Pproxy–ligand complex provides a quantitative measure of the affinity of the corresponding PT–ligand interaction. A mathematical framework for the implementation of the method in the case of monovalent (single binding site) Pproxy and monovalent and multivalent (multiple equivalent and independent binding sites) PT is described. The application of the method to screen small libraries of oligosaccharides, on the basis of human histo-blood group antigens and milk oligosaccharides, against an N-terminal fragment of the family 51 carbohydrate-binding module, a fucose-binding lectin from Ralstonia solanacearum, and human norovirus VA387 P particle (24-mer of the protruding domain of the capsid protein), serves to demonstrate the reliability and versatility of the assay.

  • Date created
    2016
  • Subjects / Keywords
  • Type of Item
    Article (Published)
  • DOI
    https://doi.org/10.7939/R30K26Q49
  • License
    © 2016 Han, L., Shams-Ud-Doha, K., Kitova, E. N., & Klassen, J. S. This version of this article is open access and can be downloaded and shared. The original author(s) and source must be cited.
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  • Citation for previous publication
    • Han, L., Shams-Ud-Doha, K., Kitova, E. N., & Klassen, J. S. (2016). Screening oligosaccharide libraries against lectins using the proxy protein electrospray ionization mass spectrometry assay. Analytical Chemistry, 88(16), 8224-8231. http://doi.org/10.1021/acs.analchem.6b02044
  • Link to related item
    http://doi.org/10.1021/acs.analchem.6b02044