Extended Data Fig. 6: Cryo-EM data processing of HCN1 M305L nanodisc in the absence and presence of 1 mM propofol.
From: Propofol rescues voltage-dependent gating of HCN1 channel epilepsy mutants
a, SEC chromatograms and SDS-PAGE of HCN1 M305L nanodisc purifications in the presence and absence of propofol. The nanodiscs for HCN1 M305L holo without propofol were made using MSP1E3 while the ones for HCN1 M305L with propofol were made using MSP2N2. For gel source data, see Supplementary Fig. 1. The cryoEM processing schematics are shown for b, apo HCN1 M305L nanodisc and c, HCN1 M305L nanodisc + pfl. Micrograph scalebar represents 50 nm. d, Pore diagram comparison between holo M305L nanodisc, M305L + pfl, and holo WT detergent (PDB 5U6P9) structures using HOLE63. Red indicates regions that are smaller than a single water molecule to pass, green for a single water molecule, and blue is double the radius of a single water molecule. Both holo structures contain cAMP. e, Backbone r.m.s.d. deviations of the voltage sensing domain (S1-S4) between holo M305L, holo WT (PDB 5U6P9), and M305L HCN1 propofol structures.
