Abstract
Nucleotide binding oligomerization domain-containing protein 1 (NOD1) and NOD2 have been identified as intracellular receptors for bacterial peptidoglycan for almost two decades; however, the direct binding with their respective ligands has only been recently demonstrated due to the difficulty of achieving large quantity of proteins with high purity. Here we describe a strategy combining immunoprecipitation of GFP-tagged proteins and microscale thermophoresis (MST) for efficient one-step purification of NOD1-GFP and NOD2-GFP and easy measurement of the binding affinities of NOD1 or NOD2 with sphingosine-1-phosphate (S1P) using small amount of proteins (nM range). This method will allow the identification of novel agonists/antagonists for NOD1/2.
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Pei, G. (2022). Identification of Novel Endogenous NOD Ligands: Quantitative Analysis of Binding Affinities of NOD1 or NOD2 with Sphingosine-1-Phosphate Using Microscale Thermophoresis. In: Kufer, T.A., Kaparakis-Liaskos, M. (eds) Effector-Triggered Immunity. Methods in Molecular Biology, vol 2523. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2449-4_10
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DOI: https://doi.org/10.1007/978-1-0716-2449-4_10
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