PLoS One. 2009;4(4):e4931. Epub 2009 Apr 1.

Evaluation of Nod-like receptor (NLR) effector domain interactions.External

Wagner, R. N., Proell, M., Kufer, T. A., Schwarzenbacher, R.,
--- - Department of Molecular Biology, University of Salzburg, Salzburg, Austria.
Members of the Nod-like receptor (NLR) family recognize intracellular pathogens and recruit a variety of effector molecules, including pro-caspases and kinases, which in turn are implicated in cytokine processing and NF-kappaB activation.In order to elucidate the intricate network of NLR signaling, which is still fragmentary in molecular terms, we applied comprehensive yeast two-hybrid analysis for unbiased evaluation of physical interactions between NLRs and their adaptors (ASC, CARD8) as well as kinase RIPK2 and inflammatory caspases (C1, C2, C4, C5) under identical conditions. Our results confirmed the interaction of NOD1 and NOD2 with RIPK2, and between NLRP3 and ASC, but most importantly, our studies revealed hitherto unrecognized interactions of NOD2 with members of the NLRP subfamily. We found that NOD2 specifically and directly interacts with NLRP1, NLRP3 and NLRP12. Furthermore, we observed homodimerization of the RIPK2 CARD domains and identified residues in NOD2 critical for interaction with RIPK2.In conclusion, our work provides further evidence for the complex network of protein-protein interactions underlying NLR function.
PMID: 19337385External
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
In vitro interaction
  DD1 DD2 Reference
Family DD1 DD2 Method Species Region Expression Species Region Expression
CARD Apaf1 Link Caspase9 Yeast two-hybrid Human 1-92 Yeast Human 1-97 Yeast 19337385
CARD ASC Link ASC Yeast two-hybrid Human 105-195 Yeast Human 105-195 Yeast 19337385
CARD ASC Link CARD12 Yeast two-hybrid Human Full length Yeast Human 1-96 Yeast 19337385
CARD ASC Link Caspase1 Yeast two-hybrid Human 105-195 Yeast Human 1-91 Yeast 19337385
CARD CARD8 Link CARD8 Yeast two-hybrid Human 342-430 Yeast Human 342-430 Yeast 19337385
CARD CARD8 Link Caspase1 Yeast two-hybrid Human 342-430 Yeast Human 1-91 Yeast 19337385
CARD Caspase2 Link Caspase2 Yeast two-hybrid Human 32-121 Yeast Human 32-121 Yeast 19337385
CARD NOD1 Link RIPK2 Yeast two-hybrid Human 1-110 Yeast Human 427-527 Yeast 19337385
CARD NOD1 Link RIPK2 Yeast two-hybrid Human 16-108 Yeast Human 427-527 Yeast 19337385
CARD NOD2 Link NOD2 Yeast two-hybrid Human 1-267 Yeast Human 1-267 Yeast 19337385
CARD NOD2 Link RIPK2 Yeast two-hybrid Human 1-267 Yeast Human 427-527 Yeast 19337385
CARD RIPK2 Link RIPK2 Yeast two-hybrid Human 427-527 Yeast Human 427-527 Yeast 19337385
PYD ASC Link ASC Yeast two-hybrid Human 1-94 Yeast Human 1-94 Yeast 19337385
PYD ASC Link NLRP3 Yeast two-hybrid Human Full length Yeast Human 1-101 Yeast 19337385
(Link: click this icon to show interactions only between the two corresponding DDs)
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
Characterization
  Stoichiometry Affinity Protein-Protein interface Reference
Family DD1 DD2 Method Mutation Complex structure
CARD NOD2 Link RIPK2 Yeast two-hybrid E69K, D70K (Nod2) 19337385
(Link: click this icon to show interactions only between the two corresponding DDs)