The heteromeric cyclic nucleotide-gated channel adopts a 3A:1B stoichlometry

Cyclic nucleotide-gated (CNG) channels are crucial for visual and olfactory transductions^1-4. These channels are tetramers and in their native forms are composed of A and B subunits⁵, with a stoichiometry thought to be 2A:2B (refs 6, 7). Here we report the identification of a leucine-zipper⁸-homology domain named CLZ (for carboxy-terminal leucine zipper). This domain is present in the distal C terminus of CNG channel A subunits but is absent from B subunits, and mediates an inter-subunit interaction. With cross-linking, non-denaturing gel electrophoresis and analytical centrifugation, this CLZ domain was found to mediate a trimeric interaction. In addition, a mutant cone CNG channel A subunit with its CLZ domain replaced by a generic trimeric leucine zipper produced channels that behaved much like the wild type, but less so if replaced by a dimeric or tetrameric leucine zipper. This A-subunit-only, trimeric interaction suggests that heteromeric CNG channels actually adopt a 3A:1B stoichiometry. Biochemical analysis of the purified bovine rod CNG channel confirmed this conclusion. This revised stoichiometry provides a new foundation for understanding the structure and function of the CNG channel family.