Irritant-evoked activation and calcium modulation of the TRPA1 receptor

The TRPA1 ion channel is expressed by primary afferent nerve fibers, where it functions as a low threshold sensor for structurally diverse electrophilic irritants ranging from small volatile environmental toxicants to endogenous algogenic lipids(1). TRPA1 is also a ‘receptor-operated’ channel whose activation downstream of metabotropic receptors elicits inflammatory pain or itch, making it an attractive target for novel analgesic therapies(2). However, we lack mechanistic insight into how TRPA1 recognizes and responds to electrophiles or cytoplasmic second messengers. Here, we show that electrophiles act through a two-step process in which modification of a highly reactive cysteine (C621) promotes reorientation of a cytoplasmic loop to enhance nucleophilicity and modification of a nearby cysteine (C665), thereby stabilizing the loop in an activating configuration. These actions modulate two restrictions controlling ion permeation, including widening of the selectivity filter to enhance calcium permeability and opening of a canonical gate at the cytoplasmic end of the pore. We propose a model to explain functional coupling between electrophile action and these control points. We also characterize a calcium binding pocket that is remarkably conserved across TRP channel subtypes and accounts for all aspects of calcium-dependent TRPA1 regulation, including potentiation, desensitization, and activation by metabotropic receptors. These findings provide a structural framework for understanding how a broad-spectrum irritant receptor is controlled by endogenous and exogenous agents that elicit or exacerbate pain and itch.