DPP9 sequesters the C terminus of NLRP1 to repress inflammasome activation.
Hollingsworth LR, Sharif H, Griswold AR, Fontana P, Mintseris J, Dagbay KB, Paulo JA, Gygi SP, Bachovchin DA, Wu H
Nature. 2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi: 10.1038/s41586-021-03350-4.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 166787 | pFastBac HTB DPP9S | Insect cell expression vector for DPP9S (protein purification) |
| 166788 | pFastBac HTB DPP9L | Insect cell expression vector for DPP9L (protein purification) |
| 166789 | pDONR221 DPP9S | pDONR gateway shuttling vector with DPP9S (no stop codon) |
| 166790 | pDONR221 DPP9L | pDONR gateway shuttling vector with DPP9L (no stop codon) |
| 166791 | pcDNA3.1 LIC 6A HIS-TEV-DPP9S | Mammalian cell expression vector for His-tagged DPP9S |
| 166793 | pcDNA3.1 LIC 6A HIS-TEV-DPP8 | Mammalian cell expression vector for Flag-tagged DPP8 |
| 166794 | pcDNA3.1 LIC 6A FLAG-TEV-DPP9S | Mammalian cell expression vector for Flag-tagged DPP9S |
| 166795 | pcDNA3.1 LIC 6A FLAG-TEV-DPP8 | Mammalian cell expression vector for FLAG-tagged DPP8 |
| 166796 | pcDNA3.1 LIC 6A FLAG-TEV-DPP9S KK93EE | Mammalian cell expression vector for FLAG-tagged DPP9S. KK93EE mutant is partial loss of CARD8 binding. |
| 166797 | pcDNA3.1 LIC 6A FLAG-TEV-DPP9S RK96EE | Mammalian cell expression vector for FLAG-tagged DPP9S. RK96EE mutant is partial loss of CARD8 binding. |
| 166798 | pcDNA3.1 LIC 6A FLAG-TEV-DPP9S LL100EE | Mammalian cell expression vector for FLAG-tagged DPP9S. LL100EE mutant is loss of CARD8 and NLRP1 binding. |
| 166799 | pcDNA3.1 LIC 6A FLAG-TEV-DPP9S LL102EE | Mammalian cell expression vector for FLAG-tagged DPP9S. LL102EE mutant is partial loss of CARD8 binding. |
| 166800 | pcDNA3.1 LIC 6A FLAG-TEV-DPP9S E597R | Mammalian cell expression vector for FLAG-tagged DPP9S. E597R mutant is partial loss of CARD8 binding. |
| 166801 | pcDNA3.1 LIC 6A FLAG-TEV-DPP9S S730A | Mammalian cell expression vector for FLAG-tagged DPP9S. The S730A mutant is catalytically dead. |
| 166802 | pcDNA3.1 LIC 6A FLAG-TEV-DPP9S D808N | Mammalian cell expression vector for FLAG-tagged DPP9S. The D808N mutant is catalytically dead. |
| 166803 | pcDNA 3.1 LIC 6A NLRP1-FLAG | Mammalian cell expression vector for FLAG-tagged NLRP1. |
| 166804 | pcDNA 3.1 LIC 6A NLRP1-FLAG S1213A | Mammalian cell expression vector for FLAG-tagged NLRP1. The S1213A mutant is deficient in FIIND autoprocessing |
| 166805 | pcDNA 3.1 LIC 6A NLRP1-FLAG LL1193EE | Mammalian cell expression vector for FLAG-tagged NLRP1. The LL1193E mutant abolishes FL-NLRP1 association with DPP9 at interface I. |
| 166806 | pcDNA 3.1 LIC 6A NLRP1-FLAG P1214R | Mammalian cell expression vector for FLAG-tagged NLRP1. The P1214R patient mutant abolishes NLRP1-CT association with DPP9 at interface II. |
| 166807 | pcDNA 3.1 LIC 6A NLRP1-FLAG H1276G | Mammalian cell expression vector for FLAG-tagged NLRP1. The H1276G mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity |
| 166808 | pcDNA 3.1 LIC 6A NLRP1-FLAG K1277E | Mammalian cell expression vector for FLAG-tagged NLRP1. The K1277E mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity |
| 166809 | pcDNA 3.1 LIC 6A NLRP1-FLAG E1322R | Mammalian cell expression vector for FLAG-tagged NLRP1. The E1322R mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity |
| 166810 | pcDNA 3.1 LIC 6A NLRP1-FLAG LL1193EE P1214R | Mammalian cell expression vector for FLAG-tagged NLRP1. LL1193EE and P1214R mutations abolish DPP9 association. |
| 166811 | pcDNA 3.1 LIC 6A NLRP1-FLAG LL1193EE P1214R K1277E | Mammalian cell expression vector for FLAG-tagged NLRP1. LL1193EE and P1214R mutations abolish DPP9 association, K1277E abolishes UPA-UPA contacts |
| 166812 | pcDNA 3.1 LIC 6A NLRP1 UPA-CARD-FLAG H1276G | Mammalian cell expression vector for FLAG-tagged UPA-CARD of NLRP1. The H1276G mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity |
| 166813 | pcDNA 3.1 LIC 6A NLRP1 UPA-CARD-FLAG K1277E | Mammalian cell expression vector for FLAG-tagged UPA-CARD of NLRP1. The K1277E mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity |
| 166814 | pcDNA 3.1 LIC 6A NLRP1 UPA-CARD-FLAG E1322R | Mammalian cell expression vector for FLAG-tagged UPA-CARD of NLRP1. The E1322R mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity |
| 166815 | pDONR221 NLRP1 A54T | Gateway pDONR vector encoding NLRP1 with the patient mutation A54T (Zhong et al., 2016) (no stop codon) |
| 166816 | pDONR221 NLRP1 A59P | Gateway pDONR vector encoding NLRP1 with the patient mutation A59P (Herlin et al., 2020) (no stop codon) |
| 166817 | pDONR221 NLRP1 A66V | Gateway pDONR vector encoding NLRP1 with the patient mutation A66V (Zhong et al., 2016) (no stop codon) |
| 166818 | pDONR221 NLRP1 M77T | Gateway pDONR vector encoding NLRP1 with the patient mutation M77T (Zhong et al., 2016) (no stop codon) |
| 166819 | pDONR221 NLRP1 R726W | Gateway pDONR vector encoding NLRP1 with the patient mutation R726W (Grandemange et al., 2017) (no stop codon) |
| 166820 | pDONR221 NLRP1 T755N | Gateway pDONR vector encoding NLRP1 with the patient mutation T755N (Drutman et al., 2019) (no stop codon) |
| 166821 | pDONR221 NLRP1 delta787-843 | Gateway pDONR vector encoding NLRP1 with the patient mutation _787-843 (Zhong et al., 2016) (no stop codon) |
| 166822 | pDONR221 NLRP1 P1214R | Gateway pDONR vector encoding NLRP1 with the patient mutation P1214R (Grandemange et al., 2017) (no stop codon) |
| 166823 | pDONR221 NLRP1 | Gateway pDONR vector containing NLRP1 (no stop codon) |
| 166824 | pDONR221 NLRP1 S1213A | Gateway pDONR vector containing NLRP1 with the P1214R patient mutation (no stop codon) |
| 166825 | dTAG NLRP1 | dTAG (degron tag) NLRP1 for ligand-controlled N-terminal degradation |
| 166826 | dTAG NLRP1 P1214R | dTAG (degron tag) NLRP1 for ligand-controlled N-terminal degradation |
| 166827 | pDONR221 Ub-NLRP1 S1213 | pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate the native S1213 N-terminus (no stop) |
| 166828 | pDONR221 Ub-NLRP1 L1215 | pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate a L1215 N-terminus (no stop) |
| 166829 | pDONR221 Ub-NLRP1 L1218 | pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate a L1218 N-terminus (no stop) |
| 166830 | pDONR221 Ub-NLRP1 S1213 P1214R | pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate the native S1213 N-terminus, with the P1214R patient mutation (Grandemange et al., 2017) (no stop) |
| 166831 | pDONR221 NLRP1 FIIND (F1079M-L1373) | Gateway pDONR vector containing NLRP1 FIIND (no stop codon) |
| 166832 | pDONR221 NLRP1 FIIND (F1079M-L1373) S1213A | Gateway pDONR vector containing NLRP1 FIIND with autoprocessing deficient S1213A mutation (no stop codon) |
| 166833 | pcDNA3.1 His-NLRP1 codon-optimized | Mammalian codon-optimized NLRP1 with N-terminal and C-terminal HIS tags |
| 166834 | pcDNA3.1 NLRP1-delPYD-delCARD-TEV-GFP-FLAG codon-optimized | Expression and purification of NLRP1 (without the PYD and CARD) from mammalian cells |
| 166835 | pcDNA3 TEV-GFP-FLAG LIC cloning vector (6LD) | Modified LIC 6D backbone with a C-terminal TEV-GFP-FLAG tag. Derived from Scott Gradia's pcDNA3.1 LIC 6D by Liron David. |
| 169946 | pFastBac HTB DPP9S S730A | Purification of catalytically-dead DPP9S from insect cells |
| 169947 | pFastBac HTB DPP9S D808N | Purification of catalytically-dead DPP9S from insect cells |