A Surface Plasmon Resonance-Based Strategy to Characterize Interactions of NLR Proteins with Associated Factors

Abstract

NOD-like receptors (NLRs) are established as key regulators of the innate immune system. In recent years, an increasing number of interaction partners have been described that modulate receptor activity by direct binding. Characterizing these interactions can be challenging because these receptors tend to adopt different conformational states. We have developed a protocol that employs intracellular protein biotinylation to provide a straightforward immobilization strategy in surface plasmon resonance experiments. With this highly sensitive and label-free technique, the kinetics and affinities of NLR and co-factor interactions can be measured directly at the protein level.

Keywords: Affinity; Biotinylation; Kinetics; NOD-like receptor (NLR); Protein–protein interaction; Surface plasmon resonance (SPR).