ArticleMethods in molecular biology (Clifton, N.J.)2022
Identification of Novel Endogenous NOD Ligands: Quantitative Analysis of Binding Affinities of NOD1 or NOD2 with Sphingosine-1-Phosphate Using Microscale Thermophoresis.
Article in Methods in molecular biology (Clifton, N.J.), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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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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