| Antibodies |
| Rabbit polyclonal anti-GFP |
Thermo Fisher Scientific |
Cat#A-31852; RRID: AB_162553 |
| Rabbit polyclonal anti-Flag |
Sigma-Aldrich |
Cat#F7425; RRID: AB_439687 |
| Mouse monoclonal anti-β-actin |
Santa Cruz Biothechnology |
Cat#sc-47778 HRP; RRID: AB_2714189 |
| Mouse monoclonal anti-HCN1 |
Addgene |
Cat#180092; RRID: AB_2750810 |
| Mouse monoclonal anti-ASIC1 |
Addgene |
Cat#184198 |
| Rabbit polyclonal anti-TMEM63A |
Sigma-Aldrich |
Cat#HPA068918; RRID: AB_2686057 |
| Rabbit polyclonal anti-HA |
Abcam |
Cat#ab9110; RRID: AB_307019 |
| Bacterial and Virus Strains |
| XL10-Gold Ultracompetent E. coli |
Agilent Technologies |
Cat#200315 |
| Chemicals, Peptides, and Recombinant Proteins |
| Glutaraldehyde |
Sigma-Aldrich |
Cat#G5882 |
| Disuccinimidyl suberate |
Thermo Fisher Scientific |
Cat#A39267 |
| n-Dodecyl-β-D-Maltopyranoside (DDM) |
Anatrace |
Cat#D310-CH210 |
| lauryl maltose neopentyl glycol (LMNG) |
Anatrace |
Cat#NG310-CH210 |
| Lipofectamine 3000 |
Thermo Fisher Scientific |
Cat#L3000008 |
| Expi293 Expression Medium |
Thermo Fisher Scientific |
Cat#A1435102 |
| Polyethylenimine Max 40k |
Polysciences |
Cat#24765–1 |
| tris(2-carboxyethyl)phosphine (TCEP) |
Sigma-Aldrich |
Cat#646547 |
| Protease inhibitor cocktail |
Sigma-Aldrich |
Cat#S8830 |
| POPC:POPE:POPG lipid mix (3:1:1) |
Anatrace |
Cat#P516:P416:P616 MIX |
| 3C protease |
Sigma-Aldrich |
Cat#SAE0045 |
| MSP2N2 scaffold protein |
Sigma-Aldrich |
Cat#MSP12 |
| Critical Commercial Assays |
| Superose 6 Increase 10/300 GL |
GE Healthcare |
Cat#29–0915-96 |
| Quikchange Site-Directed Mutagenesis |
Agilent Technologies |
Cat#210515 |
| Deposited Data |
| Coordinates of human TMEM63A in Nanodiscs |
This paper |
PDB: 8EHW |
| Cryo-EM map of human TMEM63A in Nanodiscs |
This paper |
EMDB: EMD-28153 |
| Coordinates of human TMEM63B in LMNG |
This paper |
PDB: 8EHX |
| Cryo-EM map of human TMEM63B in LMNG |
This paper |
EMDB: EMD-28154 |
| Experimental Models: Cell Lines |
| Human: HEK293T cells |
ATCC |
CRL-3216 |
| Human: Expi293F suspended cells |
Thermo Fisher Scientific |
Cat#A14527 |
| Experimental Models: Organisms/Strains |
| Mouse: Tmem63bHA/HA knock-in |
Du et al. 37
|
N/A |
| Recombinant DNA |
| cDNA: Human TMEM63A |
Genscript |
Accession number NM_014698.3
|
| cDNA: Human TMEM63B |
Genscript |
Accession number NM_018426.3
|
| cDNA: Human TMEM63C |
Genscript |
Accession number NM_020431.4
|
| Plasmid: pEG BacMam |
Goehring et al. 56
|
N/A |
| Plasmid: OSCA1.2-PP-EGFP |
Jojoa-Cruz et al. 22
|
N/A |
| Software and Algorithms |
| Serial EM |
Mastronarde 57
|
https://bio3d.colorado.edu/SerialEM
|
| MotionCor2 |
Zheng et al. 58
|
https://emcore.ucsf.edu/cryoem-software
|
| CTFFIND4 |
Rohou and Grigorieff 59
|
https://grigoriefflab.umassmed.edu/ctf_estimation_ctffind_ctftilt
|
| Relion 3.0 |
Scheres 60
|
https://www3.mrc-lmb.cam.ac.uk/relion/
|
| crYOLO |
Wagner et al. 61
|
https://cryolo.readthedocs.io/en/stable/
|
| cryoSPARC V2 |
Punjani et al. 62
|
https://cryosparc.com/
|
| SBGrid |
Morin et al. 63
|
https://sbgrid.org/
|
| UCSF Chimera |
Pettersen et al. 64
|
https://www.cgl.ucsf.edu/chimera/download.html
|
| UCSF ChimeraX |
Pettersen et al. 65
|
https://www.cgl.ucsf.edu/chimerax/download.html
|
| PyMOL |
Schrodinger, LLC |
http://www.pymol.org/
|
| COOT |
Emsley et al. 66
|
https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/
|
| PHENIX |
Adams et al. 75
|
https://phenix-online.org/download/
|
| HOLE |
Smart et al. 42
|
http://www.holeprogram.org/
|
| pClamp 10 |
Molecular Devices |
https://www.moleculardevices.com/products/axon-patch-clamp-system/acquisition-and-analysis-software/pclamp-software-suite
|
| Prism 8 |
GraphPad |
https://www.graphpad.com/scientific-software/prism/
|
| Other |
| Quantifoil R1.2/1.3 400-mesh gold grid |
Electron Microscopy Sciences |
Cat#Q3100AR1.3 |
