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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.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
166787pFastBac HTB DPP9SInsect cell expression vector for DPP9S (protein purification)
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166788pFastBac HTB DPP9LInsect cell expression vector for DPP9L (protein purification)
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166789pDONR221 DPP9SpDONR gateway shuttling vector with DPP9S (no stop codon)
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166790pDONR221 DPP9LpDONR gateway shuttling vector with DPP9L (no stop codon)
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166791pcDNA3.1 LIC 6A HIS-TEV-DPP9SMammalian cell expression vector for His-tagged DPP9S
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166793pcDNA3.1 LIC 6A HIS-TEV-DPP8Mammalian cell expression vector for Flag-tagged DPP8
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166794pcDNA3.1 LIC 6A FLAG-TEV-DPP9SMammalian cell expression vector for Flag-tagged DPP9S
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166795pcDNA3.1 LIC 6A FLAG-TEV-DPP8Mammalian cell expression vector for FLAG-tagged DPP8
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166796pcDNA3.1 LIC 6A FLAG-TEV-DPP9S KK93EEMammalian cell expression vector for FLAG-tagged DPP9S. KK93EE mutant is partial loss of CARD8 binding.
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166797pcDNA3.1 LIC 6A FLAG-TEV-DPP9S RK96EEMammalian cell expression vector for FLAG-tagged DPP9S. RK96EE mutant is partial loss of CARD8 binding.
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166798pcDNA3.1 LIC 6A FLAG-TEV-DPP9S LL100EEMammalian cell expression vector for FLAG-tagged DPP9S. LL100EE mutant is loss of CARD8 and NLRP1 binding.
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166799pcDNA3.1 LIC 6A FLAG-TEV-DPP9S LL102EEMammalian cell expression vector for FLAG-tagged DPP9S. LL102EE mutant is partial loss of CARD8 binding.
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166800pcDNA3.1 LIC 6A FLAG-TEV-DPP9S E597RMammalian cell expression vector for FLAG-tagged DPP9S. E597R mutant is partial loss of CARD8 binding.
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166801pcDNA3.1 LIC 6A FLAG-TEV-DPP9S S730AMammalian cell expression vector for FLAG-tagged DPP9S. The S730A mutant is catalytically dead.
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166802pcDNA3.1 LIC 6A FLAG-TEV-DPP9S D808NMammalian cell expression vector for FLAG-tagged DPP9S. The D808N mutant is catalytically dead.
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166803pcDNA 3.1 LIC 6A NLRP1-FLAGMammalian cell expression vector for FLAG-tagged NLRP1.
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166804pcDNA 3.1 LIC 6A NLRP1-FLAG S1213AMammalian cell expression vector for FLAG-tagged NLRP1. The S1213A mutant is deficient in FIIND autoprocessing
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166805pcDNA 3.1 LIC 6A NLRP1-FLAG LL1193EEMammalian cell expression vector for FLAG-tagged NLRP1. The LL1193E mutant abolishes FL-NLRP1 association with DPP9 at interface I.
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166806pcDNA 3.1 LIC 6A NLRP1-FLAG P1214RMammalian cell expression vector for FLAG-tagged NLRP1. The P1214R patient mutant abolishes NLRP1-CT association with DPP9 at interface II.
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166807pcDNA 3.1 LIC 6A NLRP1-FLAG H1276GMammalian cell expression vector for FLAG-tagged NLRP1. The H1276G mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity
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166808pcDNA 3.1 LIC 6A NLRP1-FLAG K1277EMammalian cell expression vector for FLAG-tagged NLRP1. The K1277E mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity
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166809pcDNA 3.1 LIC 6A NLRP1-FLAG E1322RMammalian cell expression vector for FLAG-tagged NLRP1. The E1322R mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity
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166810pcDNA 3.1 LIC 6A NLRP1-FLAG LL1193EE P1214RMammalian cell expression vector for FLAG-tagged NLRP1. LL1193EE and P1214R mutations abolish DPP9 association.
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166811pcDNA 3.1 LIC 6A NLRP1-FLAG LL1193EE P1214R K1277EMammalian cell expression vector for FLAG-tagged NLRP1. LL1193EE and P1214R mutations abolish DPP9 association, K1277E abolishes UPA-UPA contacts
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166812pcDNA 3.1 LIC 6A NLRP1 UPA-CARD-FLAG H1276GMammalian cell expression vector for FLAG-tagged UPA-CARD of NLRP1. The H1276G mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity
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166813pcDNA 3.1 LIC 6A NLRP1 UPA-CARD-FLAG K1277EMammalian cell expression vector for FLAG-tagged UPA-CARD of NLRP1. The K1277E mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity
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166814pcDNA 3.1 LIC 6A NLRP1 UPA-CARD-FLAG E1322RMammalian cell expression vector for FLAG-tagged UPA-CARD of NLRP1. The E1322R mutant abolishes interface III UPA-UPA contacts, DPP9 binding, and inflammasome activity
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166815pDONR221 NLRP1 A54TGateway pDONR vector encoding NLRP1 with the patient mutation A54T (Zhong et al., 2016) (no stop codon)
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166816pDONR221 NLRP1 A59PGateway pDONR vector encoding NLRP1 with the patient mutation A59P (Herlin et al., 2020) (no stop codon)
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166817pDONR221 NLRP1 A66VGateway pDONR vector encoding NLRP1 with the patient mutation A66V (Zhong et al., 2016) (no stop codon)
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166818pDONR221 NLRP1 M77TGateway pDONR vector encoding NLRP1 with the patient mutation M77T (Zhong et al., 2016) (no stop codon)
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166819pDONR221 NLRP1 R726WGateway pDONR vector encoding NLRP1 with the patient mutation R726W (Grandemange et al., 2017) (no stop codon)
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166820pDONR221 NLRP1 T755NGateway pDONR vector encoding NLRP1 with the patient mutation T755N (Drutman et al., 2019) (no stop codon)
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166821pDONR221 NLRP1 delta787-843Gateway pDONR vector encoding NLRP1 with the patient mutation _787-843 (Zhong et al., 2016) (no stop codon)
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166822pDONR221 NLRP1 P1214RGateway pDONR vector encoding NLRP1 with the patient mutation P1214R (Grandemange et al., 2017) (no stop codon)
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166823pDONR221 NLRP1Gateway pDONR vector containing NLRP1 (no stop codon)
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166824pDONR221 NLRP1 S1213AGateway pDONR vector containing NLRP1 with the P1214R patient mutation (no stop codon)
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166825dTAG NLRP1dTAG (degron tag) NLRP1 for ligand-controlled N-terminal degradation
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166826dTAG NLRP1 P1214RdTAG (degron tag) NLRP1 for ligand-controlled N-terminal degradation
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166827pDONR221 Ub-NLRP1 S1213pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate the native S1213 N-terminus (no stop)
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166828pDONR221 Ub-NLRP1 L1215pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate a L1215 N-terminus (no stop)
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166829pDONR221 Ub-NLRP1 L1218pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate a L1218 N-terminus (no stop)
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166830pDONR221 Ub-NLRP1 S1213 P1214RpDONR221 encoding ubiquitin-tagged NLRP1-CT to generate the native S1213 N-terminus, with the P1214R patient mutation (Grandemange et al., 2017) (no stop)
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166831pDONR221 NLRP1 FIIND (F1079M-L1373)Gateway pDONR vector containing NLRP1 FIIND (no stop codon)
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166832pDONR221 NLRP1 FIIND (F1079M-L1373) S1213AGateway pDONR vector containing NLRP1 FIIND with autoprocessing deficient S1213A mutation (no stop codon)
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166833pcDNA3.1 His-NLRP1 codon-optimizedMammalian codon-optimized NLRP1 with N-terminal and C-terminal HIS tags
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166834pcDNA3.1 NLRP1-delPYD-delCARD-TEV-GFP-FLAG codon-optimizedExpression and purification of NLRP1 (without the PYD and CARD) from mammalian cells
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166835pcDNA3 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.
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169946pFastBac HTB DPP9S S730APurification of catalytically-dead DPP9S from insect cells
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169947pFastBac HTB DPP9S D808NPurification of catalytically-dead DPP9S from insect cells
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Antibodies from Article