NOD2 directly interacts with NLRP1, NLRP3, and NLRP12.
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Abstract
<p><i>Left panel</i>, Yeast two-hybrid analysis of NOD2 CARDs (residues 1β267) showed interaction with NLRP1,-3 and -12, but not with other NLRP proteins (NLRP2, -7, -10, or -11; column 1). Lam c was used as a negative control (column 2). Interestingly, NOD2 mutant E69K maintained the binding to NLRP1, -3, and -12 (column 3), whereas NOD2 D70K (column 4) as well as the NOD2 triple mutant (3xmut, column 5) did not. An unrelated mutation not located within the prospective interaction interface had no effect (column 6). <i>Right panel</i>, Physical interaction of NOD2 and NLRP3 in human cells. Western analysis of lysates (IN) and immunoprecipitated complexes (IP) from HEK293T cells, transiently transfected with expression plasmid encoding human HA-NOD2 and FLAG-NLRP3. NLRP3 was immunoprecipitated from cell lysates using a FLAG-epitope specific antibody. Proteins were detected using anti-HA and anti-FLAG antibodies, respectively. As negative control, proteins were immunoprecipitated with FLAG-epitope specific antibody from lysates of HEK293T cells transiently transfected with HA-NOD2, but not FLAG-NLRP3.</p