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Synthetic ALPK1-TIFA inducer
InvivoGen has synthesized and purified ADP-L-glycero-b-D-manno-heptose (ADP-Heptose), an intermediary sugar in the biosynthesis of lipopolysaccharide (LPS), an essential component of the outer membrane of Gram‑negative bacteria.
ADP-Heptose has been identified as a potent pathogen-associated molecular pattern (PAMP) of Gram-negative bacteria (e.g. Helicobacter pylori and Shigella flexneri) that binds to the cytosolic pattern recognition receptor (PRR) ALPK1 [1-3]. By binding to ALPK1, ADP-Heptose triggers the oligomerization of TIFA (TRAF-interacting protein with FHA domain-containing protein A) and TRAF6 (TNF receptor-associated factor 6). This ultimately results in the activation of NF-κB and a strong pro-inflammatory response through the secretion of IL-8 .
ADP-Heptose is directly delivered to host cells (i.e. human embryonic kidney cells, intestinal epithelial cells, gastric cells, and cervical cancer cells) when bacteria are endocytosed. Additionally, it can be delivered via type III and type IV bacterial secretion systems (e.g. Yersinia pseudotuberculosis and H. pylori, respectively) [3,4]. Furthermore, extracellular ADP-Heptose can freely penetrate the host cell membrane and access the host cytoplasm. Thus, extracellular bacteria that do not encode these secretion systems (e.g. Neisseria meningitidis) are also able to activate the ALPK1-TIFA signaling axis [1, 3].
Features of ADP-Heptose:
- Potent LPS-intermediary metabolite produced by all Gram-negative bacteria
- Activates the cytosolic ALPK1-TIFA signaling pathway
- Easily penetrates the cell wall for delivery to the host cell cytoplasm
InvivoGen's ADP-Heptose is of the highest quality, guaranteed free of bacterial contamination, and has been functionally validated on HEK-Blue™ Null1-v cells. Additionally, to foster research into ADP-Heptose-dependent signaling, InvivoGen provides HEK-Blue™ KO-ALPK1 and HEK-Blue™ KO-TIFA cells.
1. Pfannkuch, L. et al. 2019. ADP heptose, a novel pathogen-associated molecular pattern identified in Helicobacter pylori. FASEB J, fj201802555R.
2. Garcia-Weber, D. et al. 2018. ADP-heptose is a newly identified pathogen-associated molecular pattern of Shigella flexneri. EMBO Rep 19
3. Zhou, P. et al. 2018. Alpha-kinase 1 is a cytosolic innate immune receptor for bacterial ADP-heptose. Nature 561, 122-126.
4. Zimmermann, S. et al. 2017. ALPK1- and TIFA-Dependent Innate Immune Response Triggered by the Helicobacter pylori Type IV Secretion System. Cell Rep 20, 2384-2395.
ADP-Heptose induced NF-κB response. HEK-Blue™ Null1-v, HEK-Blue™ KO‑ALPK1, and HEK-Blue™ KO‑TIFA cells were incubated with increasing concentrations of ADP‑Heptose (0 - 30 µg/ml). After overnight incubation, the NF-κB response was assessed by measuring the activity of SEAP in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent. Data are presented as optical density (OD) at 630 nm (mean ± SEM). The EC50 value is indicated (± std error).
Formula: C17H27N5O16P2 . Et3NH (stable form)
Molecular weight: 720.21 g/mol (stable form)
Solubility: 10 mg/ml in H2O
Working concentration: 0.01 - 30 µg/ml
- Purity >95% (UHPLC)
- Activation of the ALPK1/TIFA signaling pathway has been confirmed using a cellular assay.
- Absence of bacterial contamination (i.e. endotoxins) has been confirmed using a kinetic chromogenic LAL assay, with an endotoxin level <1 EU/mg
- 250 µg ADP-Heptose (provided as a dried powder)
ADP-Heptose is shipped at room temperature.
Store at -20°C. Upon resuspension, prepare aliquots of ADP-Heptose and store at -20°C.
Resuspended product is stable 3 months at -20°C.
Avoid repeated freeze-thaw cycles.Back to the top
Chemical structure of ADP-Heptose:
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