KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| DPP9 rabbit polyclonal Ab | Abcam | ab42080 |
| FLAG® M2 monoclonal Ab | Sigma | F3165 |
| GAPDH rabbit monoclonal Ab | Cell Signaling Technology | 14C10 |
| CARD8 C-terminal rabbit polyclonal Ab | Abcam | ab24186 |
| V5 rabbit polyclonal Ab | Abcam | ab9116 |
| CASP1 rabbit polyclonal Ab | Cell Signaling Technology | 2225 |
| GSDMD rabbit polyclonal Ab | Novus | NBP2-33422 |
| MYC tag rabbit monoclonal Ab | Cell Signaling Technology | 2278 |
| HA tag rabbit monoclonal Ab | Cell Signaling Technology | 3724 |
| Actin rabbit monoclonal Ab | Abcam | ab179467 |
| IRDye 800CW anti-rabbit | LI-COR | 925-32211 |
| IRDye 800CW anti-mouse | LI-COR | 925-32210 |
| IRDye 680CW anti-rabbit | LI-COR | 925-68073 |
| IRDye 680CW anti-mouse | LI-COR | 925-68072 |
| IRDye 680RD streptavidin | LI-COR | 926-68079 |
| Experimental Models: Cell Lines | ||
| Expi293F cells | ThermoFisher | A14528 |
| HEK 293T cells | ATCC | CRL-3216 |
| CASP1/GSDMD HEK 293T Cells | Ball et al., 2020 | N/A |
| DPP8−/−DPP9−/− HEK 293T Cells | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | N/A |
| DPP9−/− HEK 293T Cells | Griswold et al., 2019 | N/A |
| Sf9 Cells | ThermoFisher (Gibco) | B82501 |
| THP-1 CARD8−/− cells | Johnson* and Taabazuing* et al. 2018 | N/A |
| THP-1 CARD8−/− + pInducer20 CARD8 cells | This paper | N/A |
| THP-1 CARD8−/− + pInducer20 CARD8 S297A cells | This paper | N/A |
| THP-1 CARD8−/− + pInducer20 CARD8 E274R cells | This paper | N/A |
| THP-1 CARD8−/− + pInducer20 CARD8 L368G cells | This paper | N/A |
| THP-1 CARD8−/− + pInducer20 CARD8 F370G cells | This paper | N/A |
| THP-1 CARD8−/− + pInducer20 CARD8 R394E cells | This paper | N/A |
| THP-1 CARD8−/− + pInducer20 CARD8 F405G cells | This paper | N/A |
| THP-1 CARD8−/− + N-dTAG-CARD8 ZUC cells | This paper | N/A |
| THP-1 CARD8−/− + pInducer20 CARD8 S297A + N-dTAG-CARD8 ZUC cells | This paper | N/A |
| Chemicals, Peptides, and Recombinant proteins | ||
| TCEP-HCL | GoldBio | TCEP1 |
| DTT | GoldBio | DTT10 |
| Trizma® base | Millipore Sigma | T1503 |
| HEPES | Millipore Sigma | H3375 |
| PBS, pH 7.4 | Millipore Sigma | P3813 |
| Sodium Chloride | Millipore Sigma | S5886 |
| Glutaraldehyde solution | Millipore Sigma | G5882 |
| 4x Laemmli Sample Buffer | BIO-RAD | 1610747 |
| 45% Glucose, sterile-filtered | Millipore Sigma | G8769 |
| Sodium valproic acid | Millipore Sigma | P4543 |
| Val-boroPro (talabostat) mesylate | Cayman | 29007 |
| dTAG-13 | Tocris | 6605 |
| Fluorophosphonate PEG23 biotin (FP-biotin) | Griswold et al., 2019. | N/A |
| bestatin methyl ester (Me-Bs) | Sigma | 200485 |
| Compound 8J | Van Goethem, S. et al. | N/A |
| Polyethylenimine | Polysciences | 23966-1 |
| Lipofectamine2000 | Invitrogen | 12566014 |
| Fugene HD | Promega | E2311 |
| Cellfectin II | ThermoFisher | 10362100 |
| TEV protease | Hao Wu Lab | N/A |
| HRV 3C protease | Hao Wu Lab | N/A |
| 3X FLAG® peptide | Sigma | F4799 |
| Uranyl formate hydrate | Electron Microscopy Sciences | 22450 |
| Software and Algorithms | ||
| cryoSPARC v2.15.0 | Punjani et al. 2017, Punjani et al., 2019 | https://cryosparc.com/ |
| RELION v3.08 and 3.1 | Scheres and Chen, 2012 Zivanov*, Nakane*, and Forsberg* et al., 2019 | https://www3.mrc-lmb.cam.ac.uk/relion//index.php?title=Main_Page |
| crYOLO | Wagner et al., 2019. | https://sphire.mpg.de/wiki/doku.php?id=pipeline:window:cryolo |
| SerialEM | Mastronarde, D.N. 2005. | https://bio3d.colorado.edu/SerialEM/ |
| Leginon | Suloway et al., 2005 | https://emg.nysbc.org/redmine/projects/leginon/wiki/Leginon_Homepage |
| EPU | ThermoFisher | https://www.fei.com/software/epu-automated-single-particles-software-for-life-sciences/ |
| Gctf | Zhang, K. 2016 | https://www.mrc-lmb.cam.ac.uk/kzhang/ |
| CTFFIND4 | Rough and Grigorieff, 2015 | https://grigoriefflab.umassmed.edu/ctffind4 |
| MotionCor2 | Zheng et al., 2017 | https://emcore.ucsf.edu/ucsf-motioncor2 |
| GemSPOT | Robertson et al., 2020 | https://github.com/mjrober101/GemSpot |
| Maestro | Schrodinger | https://www.schrodinger.com/maestro |
| Jaguar | Schrodinger | https://www.schrodinger.com/jaguar |
| Prime | Schrodinger | https://www.schrodinger.com/prime |
| Glide | Schrodinger | https://www.schrodinger.com/glide |
| Phenix | Liebschner et al. 2019 | https://www.phenix-online.org |
| Coot | Emsley et al., 2010 | https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/ |
| ResMAP | Kucukelbir et al. 2014 | http://resmap.sourceforge.net |
| UCSF Chimera | Goddard et al., 2007 | https://www.rbvi.ucsf.edu/chimera/ |
| UCSF ChimeraX | Goddard et al., 2018 | https://www.rbvi.ucsf.edu/chimerax/ |
| Pymol | Schrodinger | https://pymol.org/2/ |
| DALI | Holm 2019 | http://ekhidna2.biocenter.helsinki.fi/dali/ |
| ESPript v3.0 | Robert and Gouet 2014 | http://espript.ibcp.fr/ESPript/ESPript/ |
| Graphpad prism v7 | Graphpad | https://www.graphpad.com/scientific-software/prism/ |
| BioRender | BioRender | https://biorender.com/ |
| SBgrid | Morin et al,, 2013 | https://sbgrid.org/ |
| Deposited Data | ||
| CryoEM maps | This paper | CARD8-DPP9: EMD-22367 CARD8-DPP9-VbP: EMD-22402 CARD8-S297A-DPP9: EMD-22974 |
| Atomic models | This paper | CARD8-DPP9 PDB ID: 7JKQ CARD8-DPP9-VbP PDB ID: 7JN7 |
| Raw EM data | This paper | CARD8-DPP9: EMPIAR-10597 CARD8-DPP9-VbP: EMPIAR-10596 CARD8-S297A-DPP9: EMPIAR-10600 |
| Extended protocols | This paper | https://www.protocols.io/groups/hao-wu-lab |
| Pymol sessions, additional data | This paper | https://osf.io/x7dv8/ |
| Vectors and plasmids | ||
| pFastBac HTB | ThermoFisher | 10584027 |
| pFastBac LIC 4B | Scott Gradia | Addgene #30115 |
| pcDNA3.1 LIC 6A vector | Scott Gradia | Addgene #30124 |
| pcDNA3.1 LIC 6D vector | Scott Gradia | Addgene #30127 |
| pInducer20 CARD8-HA PAM | This paper | Addgene #169982 |
| pcDNA3.1-TEV-GFP-FLAG LIC cloning vector (6LD) | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | Addgene #166835 |
| pcDNA 3.1 His-TEV-CARD8 | This paper | By request |
| pcDNA3.1 CARD8-TEV-GFP-FLAG | This paper | Addgene #169991 |
| pcDNA3.1 CARD8-TEV-GFP-FLAG S297A | This paper | Addgene #169992 |
| pInducer20 CARD8-HA PAM | This paper | Addgene #169982 |
| pInducer20 CARD8-HA PAM S297A | This paper | Addgene #169983 |
| pInducer20 CARD8-HA PAM E274R | This paper | Addgene #169984 |
| pInducer20 CARD8-HA PAM L368G | This paper | Addgene #169985 |
| pInducer20 CARD8-HA PAM F370G | This paper | Addgene #169986 |
| pInducer20 CARD8-HA PAM R394E | This paper | Addgene #169987 |
| pInducer20 CARD8-HA PAM F405G | This paper | Addgene #169988 |
| pcDNA3.1 Ub-S297 CARD8 WT | This paper (Genscript) | Addgene #169958 |
| pcDNA3.1 Ub-S297 CARD8 L368G | This paper (Genscript) | Addgene #169959 |
| pcDNA3.1 Ub-S297 CARD8 F370G | This paper (Genscript) | Addgene #169960 |
| pcDNA3.1 Ub-S297 CARD8 R394E | This paper (Genscript) | Addgene #169961 |
| pfastBac HTB His-TEV-DPP9S | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | Addgene #166787 |
| pFastBac HTB DPP9S S730A | This paper | Addgene #169946 |
| pFastBac HTB DPP9S D808N | This paper | Addgene #169947 |
| pcDNA3.1 LIC 6A Flag-TEV-DPP9S | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | Addgene #166794 |
| pcDNA3.1 LIC 6A Flag-TEV-DPP9S S730A | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | Addgene #166801 |
| pcDNA3.1 LIC 6A Flag-TEV-DPP9S KK93EE | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | Addgene #166796 |
| pcDNA3.1 LIC 6A Flag-TEV-DPP9S RK96EE | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | Addgene #166797 |
| pcDNA3.1 LIC 6A Flag-TEV-DPP9S LL100EE | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | Addgene #166798 |
| pcDNA3.1 LIC 6A Flag-TEV-DPP9S LL102EE | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | Addgene #166799 |
| pcDNA3.1 LIC 6A Flag-TEV-DPP9S E597R | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | Addgene #166800 |
| pcDNA3.1 LIC 6A CARD8 Ub-S297-FLAG F405G | This paper | Addgene #170119 |
| pcDNA3.1 LIC 6A CARD8-FLAG | This paper | Addgene #169948 |
| pcDNA3.1 LIC 6A CARD8-FLAG S297A | This paper | Addgene #169949 |
| pcDNA3.1 LIC 6A CARD8-FLAG E274R | This paper | Addgene #169950 |
| pcDNA3.1 LIC 6A CARD8-FLAG E274R + S297A | This paper | Addgene #169951 |
| pcDNA3.1 LIC 6A CARD8-FLAG L368G | This paper | Addgene #169952 |
| pcDNA3.1 LIC 6A CARD8-FLAG F370G | This paper | Addgene #169953 |
| pcDNA3.1 LIC 6A CARD8-FLAG R394E | This paper | Addgene #169954 |
| pcDNA3.1 LIC 6A CARD8-FLAG R394E + S297A | This paper | Addgene #169955 |
| pcDNA3.1 LIC 6A CARD8-FLAG F405G | This paper | Addgene #169956 |
| pcDNA3.1 LIC 6A CARD8-FLAG F405G + S297A | This paper | Addgene #169957 |
| pcDNA3.1 LIC 6A CARD8 Ub-S297-FLAG | This paper | Addgene #169958 |
| pcDNA3.1 LIC 6A CARD8 Ub-S297-FLAG L368G | This paper | Addgene #169959 |
| pcDNA3.1 LIC 6A CARD8 Ub-S297-FLAG F370G | This paper | Addgene #169960 |
| pcDNA3.1 LIC 6A CARD8 Ub-S297-FLAG R394E | This paper | Addgene #169961 |
| pLEX305-N-dTAG-CARD8 | This paper | Addgene #169989 |
| pLEX305-N-dTAG-CARD8-ZUC | This paper | Addgene #169990 |
| pLEX307 Ub-S297 CARD8 WT-V5 | This paper | By request |
| pLEX307 NLRP1 S1213A MYC | This paper | By request |
| pLEX307 Ub-S1213 NLRP1-V5 | This paper | By request |
| pDONR221 CARD8 | This paper | Addgene #169962 |
| pDONR221 CARD8 FIIND | This paper | Addgene #169963 |
| pDONR221 CARD8 ZUC | This paper | Addgene #169964 |
| pDONR221 CARD8 Ub-S297 | This paper | Addgene #169965 |
| pDONR221 CARD8 FIIND S297A | This paper | Addgene #169966 |
| pDONR221 CARD8 FIIND E274R | This paper | Addgene #169967 |
| pDONR221 CARD8 FIIND E274R + S297A | This paper | Addgene #169968 |
| pDONR221 CARD8 FIIND L368G | This paper | Addgene #169969 |
| pDONR221 CARD8 FIIND L370G | This paper | Addgene #169970 |
| pDONR221 CARD8 FIIND R394E | This paper | Addgene #169971 |
| pDONR221 CARD8 FIIND R394E + S297A | This paper | Addgene #169972 |
| pDONR221 CARD8 FIIND F405G | This paper | Addgene #169973 |
| pDONR221 CARD8 FIIND F405G + S297A | This paper | Addgene #169974 |
| pDONR221 CARD8 PAM | This paper | Addgene #169975 |
| pDONR221 CARD8 PAM S297A | This paper | Addgene #169976 |
| pDONR221 CARD8 PAM E274R | This paper | Addgene #169977 |
| pDONR221 CARD8 PAM L368G | This paper | Addgene #169978 |
| pDONR221 CARD8 PAM F370G | This paper | Addgene #169979 |
| pDONR221 CARD8 PAM R394E | This paper | Addgene #169980 |
| pDONR221 CARD8 PAM F405G | This paper | Addgene #169981 |
| pInducer20 CARD8-HA PAM | This paper | Addgene #169982 |
| pLEX307 NLRP1 FIIND S1213A-FLAG | Hollingsworth*, Sharif*, and Griswold* et al., 2020 | By request |
| Other | ||
| Anti-FLAG M2 affinity gel | Millipore Sigma | A2220 |
| Econo-Pac® Chromatography Column | BIO-RAD | 7321010 |
| Amicon Ultra 50 kDa cutoff, 15 mL | Millipore Sigma | UFC905096 |
| Amicon Ultra 100 kDa cutoff, 15 mL | Millipore Sigma | UFC910096 |
| Amicon ultra 10 kDa cutoff, 0.5 mL | Millipore Sigma | UFC5010BK |
| Amicon ultra 50 kDa cutoff, 0.5 mL | Millipore Sigma | UFC5050BK |
| Amicon ultra 100 kDa cutoff, 0.5 mL | Millipore Sigma | UFC5100BK |
| Slide-A-Lyzer, 10 kDa cutoff, 2 mL | Millipore Sigma | 88404 |
| MonoQ 5/50 GL column | Cytiva | GE17-5166-01 |
| Superdex 200 increase 10/300 GL column | Cytiva | 28990944 |
| Superose 6 increase 10/300 GL column | Cytiva | 29091596 |
| 4–15% Mini-PROTEAN® TGX™ Precast Protein Gels | BIO-RAD | 4561086 |
| iBlot™ 2 Transfer Stacks, PVDF | Invitrogen | IB24002 |
| iBlot™ 2 Transfer Stacks, nitrocellulose | Invitrogen | IB23001 |
| NuPAGE™ 4–12% Bis-Tris Midi Protein Gels | Invitrogen | WG1403A |
| Odyssey® Blocking Buffer (PBS) | LI-COR | 927-40010 |
| DC Protein Assay kit | BIO-RAD | 5000112 |
| Quantifoil 1.2/1.3 CU200 cryo-EM grids | Electron Microscopy Sciences | Q2100CR1.3 |
| Quantifoil 1.2/1.3 AU400 cryo-EM grids | Electron Microscopy Sciences | Q4100AR1.3 |
| C-Flat-1.2/1.3-4C | Electron Microscopy Sciences | CF413-50 |
| Formvar/Carbon 400 Mesh, Cu grids | Electron Microscopy Sciences | FCF400-Cu |
| CyQUANT™ LDH Cytotoxicity Assay Kit | ThermoFisher | C20300 |