Distinct functional open states of the polymodal nociceptor TRPV1

The polymodal nociceptor TRPV1 integrates diverse physical and chemical stimuli, yet whether these inputs drive a common activated state remains unresolved. Here, we show that TRPV1 activators stabilize functionally distinct open states that yield discrete single-channel conductance levels. These functional states appear to arise from distinct open-pore conformations, as evidenced by anomalous mole fraction effect measurements. Structurally, stimulus-specific changes in pore conformation were directly observed using site-directed fluorescence recordings, consistent with Rosetta-guided structural models. Notably, coactivation by two stimuli markedly slows channel opening rather than producing synergistic acceleration, suggesting potential conformational incompatibility across subunits. Together, these findings define TRPV1 gating as a stimulus-partitioned, multistate process and highlight opportunities for selective modulation of TRPV1 in pain management.