DPP9’s Enzymatic Activity and Not Its Binding to CARD8 Inhibits Inflammasome Activation

Griswold, Andrew R.; Ball, Daniel P.; Bhattacharjee, Abir; Chui, Ashley J.; Rao, Sahana D.; Taabazuing, Cornelius Y.; Bachovchin, Daniel A.

doi:10.1021/acschembio.9b00462

Cited by 59 publications

(124 citation statements)

References 31 publications

Supporting

Mentioning

109

Contrasting

Order By: Relevance

“…4d, e). These results are broadly consistent with previous results demonstrating that hDPP9 S759A failed to fully rescue hNLRP1 activation in hDPP9-deficient 293T cells 8 . DPP8/9 inhibitors like VbP have been shown to bind to the active site of hDPP9, which overlaps with the UPA-binding site.…”

Section: Role Of the Dpp9 Catalytic Activity And Dpp9-fiind Interactisupporting

confidence: 93%

“…To probe the mechanism of NLRP1 inhibition by DPP9, we co-expressed fulllength rNLRP1 and rDPP9 in insect cells and purified them using affinity chromatography. Supporting previous studies 3,8,9 , gel filtration analysis indicated that the two proteins formed a stable complex (Extended Data Fig. 2a).…”

Section: Architecture Of the 2:1 Rnlrp1-rdpp9 Complexsupporting

confidence: 84%

“…Notably, the amino acids of this ZU5interacting site are missing in DPP4 but highly conserved in DPP8 (Extended Data Fig. 4c), explaining why DPP8 but not DPP4 interacts with and inhibits Nlrp1b 8,23 . In addition, the short α-helix in the FIIND α-β6 loop packs against other two loops of rDPP9.…”

Section: Distinct Binding Modes Of the Two Bound Rnlrp1 Moleculesmentioning

confidence: 99%

“…Furthermore, inhibitors of class IV DPPs, such as Valine-boro-Proline (VbP), or CRISPR/Cas9mediated DPP8/9 knockout specifically activate NLRP1 and/or CARD8 (Ref. 6,8,9,13).…”

mentioning

confidence: 99%

See 3 more Smart Citations

Structural and biochemical mechanisms of NLRP1 inhibition by DPP9

Chai

Huang

Zhang

et al. 2020

Preprint

View full text Add to dashboard Cite

The nucleotide-binding domain (NBD) and leucine-rich repeat (LRR)-containing receptors (NLRs) mediate innate immunity by forming inflammasomes. Activation of the NLR protein NLRP1 requires auto-cleavage within its FIIND domain1-7. In resting cells, the dipeptidyl peptidase DPP9 interacts with NLRP1-FIIND and together with a related enzyme DPP8, suppresses spontaneous NLRP1 activation8,9. The mechanisms of DPP8/9-mediated NLRP1 inhibition, however, remain elusive. Here we provide structural and biochemical evidence demonstrating that rat NLRP1 (rNLRP1) interacts with rDPP9 in a stepwise manner to form a 2:1 complex. An auto-inhibited rNLRP1 molecule first interacts with rDPP9 via its ZU5 domain. This 1:1 rNLRP1-rDPP9 complex then captures the UPA domain of a second rNLRP1 molecule via a UPA-interacting site on DPP9 and dimeric UPA-UPA interactions with the first rNLRP1. The 2:1 rNLRP1-rDPP9 complex prevents NLRP1 UPA-mediated higher order oligomerization and maintains NLRP1 in the auto-inhibited state. Structure-guided biochemical and functional assays show that both NLRP1-binding and its enzymatic activity are required for DPP9 to suppress NLRP1, supporting guard-type activation of the NLR. Together, our data reveal the mechanism of DPP9-mediated inhibition of NLRP1 and shed light on activation of the NLRP1 inflammasome.

show abstract

Section: Role Of the Dpp9 Catalytic Activity And Dpp9-fiind Interactisupporting

confidence: 93%

Section: Architecture Of the 2:1 Rnlrp1-rdpp9 Complexsupporting

confidence: 84%

Section: Distinct Binding Modes Of the Two Bound Rnlrp1 Moleculesmentioning

confidence: 99%

“…Furthermore, inhibitors of class IV DPPs, such as Valine-boro-Proline (VbP), or CRISPR/Cas9mediated DPP8/9 knockout specifically activate NLRP1 and/or CARD8 (Ref. 6,8,9,13).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Structural and biochemical mechanisms of NLRP1 inhibition by DPP9

Chai

Huang

Zhang

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…Notably, DPP9 (as well as DPP8) was recently discovered to directly bind the FIINDs of both hNLRP1 and CARD8. 62,68 The catalytic activity of DPP9 was not required for these interactions, as the catalytically dead S759A mutant DPP9 still bound to both hNLRP1 and CARD8. Despite these similarities, however, the hNLRP1-DPP9 and CARD8-DPP9 interactions appear to be remarkably distinct (Table 2).…”

Section: The D Ipep Tidyl Pep Tida S E S 8 and 9 (D Pp8/9)mentioning

confidence: 99%

The NLRP1 and CARD8 inflammasomes

Taabazuing

Griswold

Bachovchin

2020

Immunological Reviews

Self Cite

136

View full text Add to dashboard Cite

Inflammasomes are multiprotein complexes that activate inflammatory cytokines and induce pyroptosis in response to intracellular danger‐associated signals. NLRP1 and CARD8 are related germline‐encoded pattern recognition receptors that form inflammasomes, but their activation mechanisms and biological purposes have not yet been fully established. Both NLRP1 and CARD8 undergo post‐translational autoproteolysis to generate two non‐covalently associated polypeptide chains. NLRP1 and CARD8 activators induce the proteasome‐mediated destruction of the N‐terminal fragment, liberating the C‐terminal fragment to form an inflammasome. Here, we review the danger‐associated stimuli that have been reported to activate NLRP1 and/or CARD8, including anthrax lethal toxin, Toxoplasma gondii, Shigella flexneri and the small molecule DPP8/9 inhibitor Val‐boroPro, focusing on recent mechanistic insights and highlighting unresolved questions. In addition, we discuss the recently identified disease‐associated mutations in NLRP1 and CARD8, the potential role that DPP9’s protein structure plays in inflammasome regulation, and the emerging link between NLRP1 and metabolism. Finally, we summarize all of this latest research and consider the possible biological purposes of these enigmatic inflammasomes.

show abstract

Vildagliptin‐Derived Dipeptidyl Peptidase 9 (DPP9) Inhibitors: Identification of a DPP8/9‐Specific Lead

et al. 2022

View full text Add to dashboard Cite

Vildagliptin is a marketed DPP4 inhibitor, used in the management of type 2 diabetes. The molecule also has notable DPP8/9 affinity, with some preference for DPP9. Therefore, we aimed to use vildagliptin as a starting point for selective DPP8/9 inhibitors, and to engineer out the parent compound's DPP4affinity. In addition, we wanted to identify substructures in the obtained molecules that allow their further optimization into inhibitors with maximal DPP9 selectivity. Various 2S-cyanopyrrolidines and isoindoline were investigated as P1 residues of vildagliptin analogs. The obtained set was expanded with derivatives bearing O-substituted, N-(3-hydroxyadamantyl)glycine moieties at the P2 position. In this way, representatives were discovered with DPP8/9 potencies comparable to the parent molecule, but with overall selectivity towards DPP4, DPP2, FAP, and PREP. Furthermore, the most promising molecules in this series have a 4-to 7-fold preference for DPP9 over DPP8. Finally, a molecular dynamics study was carried out to maximize our insight into experimental selectivity data.

show abstract

DPP9’s Enzymatic Activity and Not Its Binding to CARD8 Inhibits Inflammasome Activation

Cited by 59 publications

References 31 publications

Structural and biochemical mechanisms of NLRP1 inhibition by DPP9

Structural and biochemical mechanisms of NLRP1 inhibition by DPP9

The NLRP1 and CARD8 inflammasomes

Vildagliptin‐Derived Dipeptidyl Peptidase 9 (DPP9) Inhibitors: Identification of a DPP8/9‐Specific Lead

Contact Info

Product

Resources

About