ION CHANNEL THERMODYNAMICS STUDIED WITH TEMPERATURE JUMPS MEASURED AT THE CELL MEMBRANE

Abstract

AbstractPerturbing the temperature of a system modifies its energy landscape thus providing a ubiquitous tool to understand biological processes. Here, we developed a framework to generate sudden temperature jumps (Tjumps) and sustained temperature steps (Tsteps) to study the temperature dependence of membrane proteins under voltage-clamp, while measuring the membrane temperature. Utilizing the melanin under the Xenopus laevis oocytes membrane as a photothermal transducer, we achieved short Tjumps up to 10 ºC in less than 1.5 ms and constant Tsteps for durations up to 150 ms. We followed the temperature at the membrane with submillisecond time resolution by measuring the time-course of membrane capacitance, which is linearly related to temperature. We applied Tjumps in Kir 1.1b, which reveals a highly temperature-sensitive blockage relief and characterized the effects of Tsteps on the temperature-sensitive channels TRPM8 and TRPV1. These newly developed approaches provide a general tool to study membrane proteins thermodynamics.