Figure 5.

Calmodulin mediates the association of TRPA1 with MORs. The recombinant cytosolic Nt and Ct regions of TRPA1 were covalently attached to N-Hydroxysuccinimide (NHS)-agarose beads and then incubated with MOR in the absence and presence of 3 mM CaCl₂. In another set of assays, the TRPA1 regions were sequentially incubated with calmodulin (CaM) and MORs in the presence of 3 mM CaCl₂. At the end of the incubation, agarose-TRPA1 was recovered by several cycles of washing-resuspension and the bound proteins were detached with 2× Laemmli buffer and resolved by SDS-PAGE followed by immunodetection. Further details in the Methods. Key: Input, Proteins incubated with agarose TRPA1, MORs are immunodetected; WB, immunodetected protein in Western blot.