Picodiscs for facile protein-glycolipid interaction analysis

• Author(s) / Creator(s)

  • Klassen, John S.
  • Cairo, Christopher W.Xiong, Zi Jian
  • Li, Jun
  • Privé, Gilbert G.
  • Leney, Aneika C.
  • Kitova, Elena N.
  • Darestani, Reza Rezaei
  • Zou, Chunxia
  • Nikjah, Sanaz

• Protein interactions with glycolipids are implicated in diverse cellular processes. However, the study of protein–glycolipid complexes remains a significant experimental challenge. Here, we describe a powerful new assay that combines electrospray ionization mass spectrometry (ESI-MS) and picodiscs, which are composed of human sphingolipid activator protein saposin A and a small number of phospholipids, to display glycolipids in a lipid environment for protein–glycolipid interaction studies in aqueous solution. Time-resolved measurements of enzyme catalyzed hydrolysis of glycolipid substrates and the detection of low, moderate, and high affinity protein–glycolipid interactions serve to demonstrate the reliability and versatility of the assay.

• Date created

  2015

• Subjects / Keywords

  • Microarrays
  • Mammalian sialidases
  • Substrate recognition
  • Ligands
  • Phospholipid-bilayer nanodiscs
  • Ionization mass-spectrometry
  • GM1 ganglioside
  • Cholera-toxin binding
  • Receptor
  • Membrane-proteins

• Type of Item

  Article (Published)

• DOI

  https://doi.org/10.7939/R39S1M01W

• License

  © 2015 Leney, A. C., Darestani, R. R., Li, J., Nikjah, S., Kitova, E. N., Zou, C., Cairo, C. W., Xiong, Z. J., Prive, G. G., & 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.