Journal: Structure (London, England : 1993)

Article Title: Structure of mechanically activated ion channel OSCA2.3 reveals mobile elements in the transmembrane domain.

doi: 10.1016/j.str.2023.11.009

Figure Lengend Snippet: Figure 1. Structures of OSCA2.3 and OSCA1.2 in peptidiscs DeepEMhancer maps (A, D), side (B, E) and top (C, F) views of models of OSCA2.3 and OSCA1.2, respectively. Ex: Extracellular, In: Intracellular. NSPr (peptidisc) peptides that were resolved in the OSCA1.2 structure are colored orange in D–F. (G) Close-up view from c displaying the EICs. (H) Right: Close-up view of the inter-subunit cleft of OSCA2.3 (salmon) and OSCA1.2 (blue). Black lines represent the approximate height at which cross-sectional area was measured. Left: Representation of the shape of the cross sections and measurements of the area occupied. (I) Superposition of Gaussian-filtered unsharpened maps showing difference in overall peptidisc shape of OSCA2.3 (red) and OSCA1.2 (blue) (see also Video S1).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins PresScission Protease Cytiva Cat#27084601 Expifectamine Sf transfection reagent Life Technologies Cat#A38915 CNBr-Activated Sepharose resin Cytiva Cat#45-000-066 Bio-beads SM-2 Bio-Rad Cat#1523920 NSPr peptide (Bulk Peptidisc) Peptidisc Biotech N/A Amicon Ultra centrifugal filter 100 kDa MWCO Milipore Cat#UFC910024 Superose 6 increase 10/300 GL Cytiva Cat#29091596 n-dodecyl-b-D-maltoside (DDM) Anatrace Cat#D310LA Cholesteryl hemisuccinate (CHS) tris salt Sigma-Aldrich Cat#C6013-25G Deposited data Map for cryo-EM structure of OSCA1.2 in peptidisc This study EMDB: EMD-41043 Coordinates for cryo-EM structure of OSCA1.2 in peptidisc This study PDB: 8T56 Map for cryo-EM structure of OSCA2.3 in peptidisc This study EMDB: EMD-41044 Coordinates for cryo-EM structure of OSCA2.3 in peptidisc This study PDB: 8T57 Map for cryo-EM structure of OSCA1.2 in nanodisc Jojoa-Cruz et al., 201820 EMDB: EMD-9112 Coordinates for cryo-EM structure of OSCA1.2 in nanodisc Jojoa-Cruz et al., 201820 PDB:6MGV Experimental models: Cell lines Sf9 cells ATCC Cat#CRL-1711 HEK293F cells ATCC Cat#R79007 Recombinant DNA Plasmid: OSCA2.3-mGFP-Strep-tagII in pEG This study N/A Plasmid: OSCA1.2-EGFP in pcDNA3.1 Jojoa-Cruz et al., 201820 N/A Software and algorithms MicAssess Li et al., 202058 https://github.com/cianfrocco-lab/ Automatic-cryoEM-preprocessing cryoSPARC v2 Punjani et al., 201757 https://cryosparc.com Gctf Zhang et al., 201659 N/A Leginon Suloway et al., 200555 https://emg.nysbc.org/redmine/projects/ leginon/wiki/Leginon_Homepage RELION Scheres et al., 201260 https://github.com/3dem/relion CTFFIND4 Rohou et al., 201562 https://grigoriefflab.umassmed.edu/ctffind4 MotionCor2 Zheng et al., 201756 https://emcore.ucsf.edu/ucsf-software SIDESPLITTER Ramlaul et al., 202063 https://www.ccpem.ac.uk/download.php Local Deblur Ramirez-Aportela et al., 202064 https://github.com/I2PC/xmipp Scipion de la Rosa-Trevin et al., 201667 https://scipion.i2pc.es// DeepEMhancer Sanchez-Garcia et al., 202168 https://github.com/rsanchezgarc/ deepEMhancer SWISS-MODEL Waterhouse et al., 201869 https://swissmodel.expasy.org UCSF Chimera Pettersen et al., 200470 https://www.cgl.ucsf.edu/chimera/ (Continued on next page) e1 Structure 32, 157–167.e1–e5, February 1, 2024

Techniques: