C12-iE-DAP

NOD1 Agonist - Acylated derivative of iE-DAP

NOD1 activation with C12-iE-DAP

C12-iE-DAP is an acylated derivative of the iE-DAP dipeptide (γ-D-Glu-mDAP). iE-DAP is the minimal motif of bacteria peptidoglycan (PGN) recognized by NOD1. It is present in the PGN of Gram-negative bacilli and particular Gram-positive bacteria such as Bacillus subtilis and Listeria monocytogenes [1]. C12-iE-DAP is a highly potent activator of the cytosolic receptor NOD1.

 More details

Mode of action:

Similar to iE-DAP, C12-iE-DAP is recognized by the intracellular sensor NOD1 (CARD4), which results in a signaling cascade involving the serine/threonine RIP2 (RICK, CARDIAK) kinase and IKK, and leading to NF-κB activation and the production of inflammatory cytokines such as TNF-α and IL-6 [2].
C12-iE-DAP stimulates NOD1 at concentrations 100- to 1000-fold lower than iE-DAP. C12-iE-DAP was generated by the addition of a lauroyl (C12) group to the glutamic residue of iE-DAP. C12-iE-DAP provided by InvivoGen is chemically synthesized and tested using HEK-Blue™ NOD1. Of note, it is a mixture of Lauroyl-γ-D-Glu-D-mDAP and Lauroyl-γ-D-Glu-L-mDAP.

Key features:

• Potent NOD1 agonist
• Each lot is functionally tested

Read our review on NOD-Like Receptors

References:

1. Chamaillard M. et al., 2003. An essential role for NOD1 in host recognition of bacterial peptidoglycan containing diaminopimelic acid. Nat. Immunol.4(7):702-7.
2. Park J.H. et al., 2007. RICK/RIP2 mediates innate immune responses induced through Nod1 and Nod2 but not TLRs. J Immunol. 178(4):2380-6.

Figures

C12-iE-DAP is a potent activator of human (h)NOD1. HEK-Blue™ hNOD1 cells were incubated in HEK-Blue™ Detection medium and stimulated with increasing concentrations of C12-iE-DAP and iE-DAP. After 24h incubation, the levels of NF-κB-induced SEAP were determined by reading the optical density (OD) at 630 nm (mean ± SEM).

Specifications

Specificity: NOD1 agonist

Working concentration: 10 ng - 10 µg/ml

Synonym: Lauroyl-γ-D-glutamyl-meso-diaminopimelic acid

Formula: C24H43N3O8

Molecular weight: 501.61 g/mol

Solubility: 10 mg/ml DMSO or methanol

Quality Control:

• Activation of NOD1 has been confirmed using HEK-Blue™ NOD1 cells.
• The absence of NOD2 activity has been tested using HEK‑Blue™ NOD2 cells.
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK‑Blue™ TLR4 cells.

Contents

C12-iE-DAP is provided as a lyophilized powder

• 1 mg C12-iE-DAP
• 1.5 ml endotoxin-free water

C12-iE-DAP is shipped at room temperature.

 Upon receipt, store at -20°C. 

Details

NOD1 and NOD2

The cytosolic NOD-Like Receptors (NLRs, also known as NODs or NALP) are Nucleotide-binding Oligomerization Domain containing receptors. To date, 22 NLRs have been identified in humans and constitute a major class of intracellular pattern recognition receptors (PRRs) [1].

The founding NLR-family members NOD1 (CARD4) and NOD2 (CARD15) recognize distinct motifs of peptidoglycan (PGN), an essential constituent of the bacterial cell wall. NOD1 senses the D-γ-glutamyl-meso-DAP dipeptide (iE-DAP), which is found in PGN of all Gram-negative and certain Gram-positive bacteria [1, 2] whereas NOD2 recognizes the muramyl dipeptide (MDP) structure found in almost all bacteria. Thus NOD2 acts as a general sensor of PGN and NOD1 is involved in the recognition of a specific subset of bacteria. Both iE-DAP and MDP must be delivered intracellularly either by bacteria that invade the cell or through other cellular uptake mechanisms. Ligand-bound NOD1 and NOD2 oligomerize and signal via the serine/threonine RIP2 kinase through CARD-CARD homophilic interactions [3]. Once activated, RIP2 mediates ubiquitination of NEMO/IKKγ leading to the activation of NF-κB and the production of inflammatory cytokines. Furthermore, poly-ubiquitinated RIP2 recruits TAK1, which leads to IKK complex activation and the activation of MAPKs [4].

Genetic mutations in NOD2 are associated with Crohn’s disease, a chronic inflammatory bowel disease [5].  In addition, numerous studies have recently revealed that NOD1 and NOD2 have a close relationship with a variety of cancers via controlling proliferation, altering immunosurveillance, and interacting with tissue bacteria, including intestinal commensal intestinal microflora. Moreover, additional research into the mechanisms of NOD1 and NOD2 in cancers would shed light on the innate immunity-cancer relationship and provide intriguing targets for immunotherapy [6].

References:

1. Chamaillard M. et al., 2003. An essential role for NOD1 in host recognition of bacterial peptidoglycan containing diaminopimelic acid. Nat. Immunol. 4: 702-707.
2. Girardin S. et al., 2003. Nod1 detects a unique muropeptide from Gram-negative bacterial peptidoglycan. Science 300: 1584-1587.
3. Kobayash, K. et al., 2002. RICK/Rip2/CARDIAK mediates signalling for receptors of the innate and adaptive immune systems. Nature 416: 194-199.
4. Kobayashi K. et al., 2005. Nod2-dependent regulation of innate and adaptive immunity in the intestinal tract. Science 307: 731-734.
5. Ogura Y. et al., 2001. A frameshift mutation in NOD2 associated with susceptibility to Crohn’s disease. Nature 411: 603-606.
6. Wang D, 2022. NOD1 and NOD2 Are Potential Therapeutic Targets for Cancer Immunotherapy. Comput Intell Neurosci.;2022:2271788.

Chemical structure of C12-iE-DAP: