Temperature sensing is crucial for homeotherms, including human beings, to maintain a stable body core temperature and respond to the ambient environment. A group of exquisitely temperaturesensitive transient receptor potential channels, termed thermoTRPs, serve as cellular temperature sensors. How thermoTRPs convert thermal energy (heat) into protein conformational changes leading to channel opening remains unknown. Here we demonstrate that the pathway for temperature-dependent activation is distinct from those for ligand-and voltage-dependent activation and involves the pore turret. We found that mutant channels with an artificial pore turret sequence lose temperature sensitivity but maintain normal ligand responses. Using site-directed fluorescence recordings we observed that temperature change induces a significant rearrangement of TRPV1 pore turret that is coupled to channel opening. This movement is specifically associated to temperature-dependent activation and is not observed during ligand-and voltage-dependent channel activation. These observations suggest that the turret is part of the temperature-sensing apparatus in thermoTRP channels, and its conformational change may give rise to the large entropy that defines high temperature sensitivity.conformational change | fluorescence resonance energy transfer | temperature sensing | thermodynamics T emperature-sensitive transient receptor potential channels, or thermoTRPs, include four heat-activated channels (TRPV1-4) and two cold-activated channels (TRPM8 and TRPA1) that exhibit nicely spaced activation temperatures covering the physiological temperature range (1-4). These are expressed in dorsal root ganglion sensory neurons, keratinocytes, and other cells (1). Temperature changes cause rapid, reversible activation of thermoTRP channels in both native cells (3) and expression systems ( Fig. 1 A and B). Depolarizing currents through these nonselective, cationpermeable channels lead to the generation of action potentials (5) or the release of messenger molecules (6, 7) that encodes temperature information. Structurally, thermoTRPs resemble voltagedependent potassium channels, with four subunits surrounding a central ion permeation pore (4). Each subunit contains six transmembrane segments (S1-S6) and long intracellular N and C termini. The channel pore is formed by S6 and a P-loop that in most thermoTRPs is noticeably longer than those of KcsA and voltagegated potassium channels (8, 9). Unique TRP channel structural elements, such as TRP Box immediately after S6 and N-terminal ankyrin repeats, are found in most thermoTRPs.Despite extensive research, the channel structure bestowing high temperature sensitivity on thermoTRPs remains elusive. ThermoTRP channels are polymodal sensors responsive to a wide range of physical and chemical stimuli, such as transmembrane voltage, ligands, and pH. It has been proposed that heat might control thermoTRP activation by shifting the channel's response to these stimuli (10, 11). Alternatively, synergistic activation by mul...