Analysis of protein-heparin interactions using a portable SPR instrument

Su, Dunhao; Li, Yong; Yates, Edwin; Skidmore, Mark; Andrade De Lima, Marcelo; Fernig, David

Abstract

<jats:p>Optical biosensors such as those based on surface plasmon resonance (SPR) are a key analytical tool for understanding biomolecular interactions and function as well as the quantitative analysis of analytes in a wide variety of settings. The advent of portable SPR instruments enables analyses in the field. A critical step in method development is the passivation and functionalisation of the sensor surface. We describe the assembly of a surface of thiolated oleyl ethylene glycol/biotin oleyl ethylene glycol and its functionalisation with streptavidin and reducing end biotinylated heparin for a portable SPR instrument. Such surfaces can be batch prepared and stored. Two examples of the analysis of heparin-binding proteins are presented. The binding of fibroblast growth factor 2 and competition for the binding of a heparan sulfate sulfotransferase by a library of selectively modified heparins and suramin, which identify the selectivity of the enzyme for sulfated structures in the polysaccharide and demonstrate suramin as a competitor for the enzyme’s sugar acceptor site. Heparin functionalised surfaces should have a wide applicability, since this polysaccharide is a close structural analogue of the host cell surface polysaccharide, heparan sulfate, a receptor for many endogenous proteins and viruses.</jats:p>

Citation

Su, D., Li, Y., Yates, E., Skidmore, M., Andrade De Lima, M., & Fernig, D. (2022). Analysis of protein-heparin interactions using a portable SPR instrument. PeerJ Analytical Chemistry, 4, 1-16. https://doi.org/10.7717/peerj-achem.15

• Journal Article Type: Article
• Acceptance Date: Feb 2, 2022
• Online Publication Date: Apr 8, 2022
• Publication Date: Apr 8, 2022
• Journal: PeerJ Analytical Chemistry
• Print ISSN: 2691-6630
• Publisher: PeerJ
• Volume: 4
• Pages: 1-16
• DOI: https://doi.org/10.7717/peerj-achem.15
• Keywords: Heparin, Heparan sulfate, Self-assembled monolayer, Surface plasmon resonance, Protein-polysaccharide interaction, Binding site footprinting, Enzyme competitive inhibition, Biotinylated heparin capture
• Publisher URL: https://peerj.com/articles/achem-15/