Journal: Cell
Article Title: Structural Basis of Nav1.7 Inhibition by a Gating-Modifier Spider Toxin.
doi: 10.1016/j.cell.2018.12.018
Figure Lengend Snippet: Figure 4. ProTx2 Positions 22 and 26 Modu- late the Nav1.7 Channel (A) Pore view of the ProTx2 receptor site. Elec- trostatic surface of ProTx2 with R22 highlighted. (B) Similar to (A); ProTx2-R22E variant modeled. (C) Potency of ProTx2 and GrTx1 derivatives measured on human Nav1.7 (by 95% confidence interval IC50) using a pulse protocol that favors closed channel states. (D) Characteristics of WT and E811A, D816A, and E818A channels. Error bars represent mean ± SEM. See also Figure S6 and Table S3.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-NavAb antibody This paper 7A9 Chemicals, Peptides, and Recombinant Proteins Ham’s F-12 Thermofisher Scientific Cat# 31765035 Tet System Approved fetal bovine serum Clontech Cat# 631106 DMEM (Dulbecco’s modified Eagle’s medium) Thermofisher Scientific Cat# 10569010 GlutaMAXTM supplement Thermofisher Scientific Cat# 35050061 Accutase Innovative Cell Technologies Cat# AT104-500 CHO-S-SFM II medium Thermofisher Scientific Cat# 31033020 MTSEA Toronto Research Chemicals Cat# A609150 MTSET Toronto Research Chemicals Cat# T795903 MTSES Toronto Research Chemicals Cat# S672000 Tetrodotoxin (TTX) Thermofisher Scientific Cat# 50-753-2807 Lipofectamine 2000 Transfection Reagent Invitrogen Cat# 11668019 ESF 921 Insect Cell Culture Medium Expression Systems Cat# 96-001-01 cOmplete EDTA-free protease-inhibitor cocktail tablets Roche Cat# 11873580001 Ni-NTA agarose QIAGEN Cat# 30210 ANTI-FLAG M2 Affinity Gel Sigma-Aldrich Cat# A2220 FLAG Peptide Sigma-Aldrich Cat# F3290 GDN Anatrace Cat# GDN101 Facade-EM Avanti Polar Lipids Cat# 850522 1-palmitoyl-2-oleoyl-glycero-3phosphocholine (POPC) Avanti Polar Lipids Cat# 850457 1-palmitoyl-2-oleoyl-sn-glycero-3phosphoethanolamine (POPE) Avanti Polar Lipids Cat# 850757 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho(1’-rac-glycerol) (POPG) Avanti Polar Lipids Cat# 840457 Brain Polar Lipid Extract (Porcine) Avanti Polar Lipids Cat# 141101 Pierce Fab Preparation Kit Thermo scientific Cat# 44985 ProTx2 Alomone Labs Cat# STP-100 ProTx2 Smartox Biotechnology Cat# 07PTX002-00100 ProTx2 variants Smartox Biotechnology N/A ProTx2 (SeCys-substituted) CPC Scientific N/A GrTx1 Alomone Labs Cat# STG-250 GrTx1 Smartox Biotechnology Cat# GRX GrTx1 variants Smartox Biotechnology N/A GsAf1 Alomone Labs Cat# STG-300 GsAf2 Alomone Labs Cat# STG-350 PaTx1 Alomone Labs Cat# STP-700 Deposited Data Crystal structure of Nav1.7 VSD2-ProTx2 This paper PDB: 6N4I Nav1.7 VSD2-ProTx2 (activated state) atomic model and density map This paper PDB: 6N4Q EMDB: EMD-0341 Nav1.7 VSD2-ProTx2 (deactivated state) atomic model and density map This paper PDB: 6N4R EMDB: EMD-0342 (Continued on next page) Cell 176, 1–14.e1–e6, February 7, 2019 e1
Techniques: Variant Assay
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