Study of PDZ-Peptide and PDZ-Lipid Interactions by Surface Plasmon Resonance/BIAcore

Methods Mol Biol. 2021:2256:75-87. doi: 10.1007/978-1-0716-1166-1_5.

Abstract

Surface plasmon resonance (SPR)/BIAcore technology enables the characterization of molecular interactions, including determination of affinities and kinetics. In BIAcore, one of the interaction partners (the ligand) is immobilized on a chip and the other (the analyte) is provided in solution. BIAcore allows to study association and dissociation rates in real time without the use of labeling. BIAcore can be applied to molecular interactions involving small compounds and biological macromolecules such as proteins, lipids, nucleic acids, or carbohydrates. Here we describe protocols for the measurements of PDZ domain-peptide (oriented biotinylated peptides), PDZ domain-liposomes (lipid membranes), and PDZ-lipid-peptide tripartite interactions.

Keywords: BIAcore; L1 sensor chip; Lipid interaction; Lipids; Liposomes; PDZ; Peptide interaction; SA sensor chip.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques / methods*
  • Humans
  • Kinetics
  • Ligands
  • Liposomes / metabolism*
  • Membrane Lipids / metabolism*
  • Membrane Proteins / metabolism*
  • PDZ Domains*
  • Peptide Fragments / metabolism*
  • Protein Binding
  • Surface Plasmon Resonance / methods*

Substances

  • Ligands
  • Liposomes
  • Membrane Lipids
  • Membrane Proteins
  • Peptide Fragments