Source data for paper "Voltage Sensor Conformations Induced by LQTS-associated Mutations in hERG Potassium Channels"

In this study, we employed a combination of techniques including nonsense suppression-mediated incorporation of noncanonical amino acids for site-specific fluorescence labeling, FLIM-based transition metal Förster resonance energy transfer (FLIM-tmFRET), and MD simulations to examine the impact of a LQTS-associated mutation on conformational rearrangements of hERG voltage sensors in transfected cell lines. We applied a dual stop-codon suppression strategy relying on a small fluorescent noncanonical amino acid to discern and dissect the conformational heterogeneity of voltage sensors by analyzing the frequency-domain phasor plot function of FLIM-tmFRET. This methodology has significant potential for broad applications, offering an effective approach to quantitatively assess the movement, states, dynamics, and energetics of voltage sensors in ion channels.