{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T21:23:53Z","timestamp":1761513833348,"version":"3.37.3"},"reference-count":21,"publisher":"Wiley","license":[{"start":{"date-parts":[[2016,1,1]],"date-time":"2016-01-01T00:00:00Z","timestamp":1451606400000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Scientific World Journal"],"published-print":{"date-parts":[[2016]]},"abstract":"<jats:p>Nucleotide-binding oligomerization domain-containing protein (Nod) 2 is an intracellular pattern recognition receptor, which recognizes muramyl dipeptide (N-Acetylmuramyl-L-Alanyl-D-Isoglutamine: MDP), a bacterial peptidoglycan component, and makes a NF-<jats:italic>\u03ba<\/jats:italic>B-activating complex called nodosome with adaptor protein RICK (RIP2\/RIPK2). Nod2 mutants are associated with the autoinflammatory diseases, Blau syndrome (BS)\/early-onset sarcoidosis (EOS). For drug discovery of BS\/EOS, we tried to develop Nod2-nodosome in a cell-free system. FLAG-tagged RICK, biotinylated-Nod2, and BS\/EOS-associated Nod2 mutants were synthesized, and proximity signals between FLAG-tagged and biotinylated proteins were detected by amplified luminescent proximity homogeneous assay (ALPHA). Upon incubation with MDP, the ALPHA signal of interaction between Nod2-WT and RICK was increased in a dose-dependent manner. The ALPHA signal of interaction between RICK and the BS\/EOS-associated Nod2 mutants was more significantly increased than Nod2-WT. Notably, the ALPHA signal between Nod2-WT and RICK was increased upon incubation with MDP, but not when incubated with the same concentrations, L-alanine, D-isoglutamic acid, or the MDP-D-isoform. Thus, we successfully developed Nod2-nodosome in a cell-free system reflecting its function in vivo, and it can be useful for screening Nod2-nodosome-targeted therapeutic molecules for BS\/EOS and granulomatous inflammatory diseases.<\/jats:p>","DOI":"10.1155\/2016\/2597376","type":"journal-article","created":{"date-parts":[[2016,6,15]],"date-time":"2016-06-15T21:02:24Z","timestamp":1466024544000},"page":"1-7","source":"Crossref","is-referenced-by-count":9,"title":["Nod2-Nodosome in a Cell-Free System: Implications in Pathogenesis and Drug Discovery for Blau Syndrome and Early-Onset Sarcoidosis"],"prefix":"10.1155","volume":"2016","author":[{"given":"Tomoyuki","family":"Iwasaki","sequence":"first","affiliation":[{"name":"Department of Pathology, Ehime University Proteo-Science Center and Graduate School of Medicine, Shitsukawa 454, Toon, Ehime 791-0295, 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