Synthesis and validation of click-modified NOD1/2 agonists

• Published in: Innate immunity (London, England) Vol. 29; no. 8; pp. 186 - 200
• Main Authors: Bharadwaj, Ravi, Anonick, Madison V., Jaiswal, Swati, Mashayekh, Siavash, Brown, Ashley, Wodzanowski, Kimberly A., Okuda, Kendi, Silverman, Neal, Grimes, Catherine L.
• Format: Journal Article
• Language: English
• Published: London, England SAGE Publications 01.11.2023; SAGE PUBLICATIONS, INC; SAGE Publishing
• Subjects: Agonists; Cytosol; Diaminopimelic Acid - metabolism; Nod1 protein; Nod1 Signaling Adaptor Protein - metabolism; NOD2 protein; Nod2 Signaling Adaptor Protein - genetics; Nod2 Signaling Adaptor Protein - metabolism; Original; Peptidoglycans; Receptor Protein-Tyrosine Kinases - metabolism; and NOD2; alkyne-modification; MDP; NOD1; muropeptide; iE-DAP; Click-chemistry
• ISSN: 1753-4259; 1753-4267
• DOI: 10.1177/17534259231207198

NOD1 and NOD2 sense small bacterial peptidoglycan fragments, often called muropeptides, that access the cytosol. These muropeptides include iE-DAP and MDP, the minimal agonists for NOD1 and NOD2, respectively. Here, we synthesized and validated alkyne-modified muropeptides, iE-DAP-Alk and MDP-Alk, for use in click-chemistry reactions. While it has long been known that many cell types respond to extracellular exposure to muropeptides, it is unclear how these innate immune activators access their cytosolic innate immune receptors, NOD1 and NOD2. The subcellular trafficking and transport mechanisms by which muropeptides access these cytosolic innate immune receptors are a major gap in our understanding of these critical host responses. The click-chemistry-enabled agonists developed here will be particularly powerful to decipher the underlying cell biology and biochemistry of NOD1 and NOD2 innate immune sensing.