Structural architecture of the human NALCN channelosome

Depolarizing sodium (Na+) leak currents carried by the NALCN channel regulate the resting membrane potential of many neurons to modulate respiration, circadian rhythm, locomotion and pain sensitivity1–8. NALCN requires FAM155A, UNC79 and UNC80 to function, but the role of these auxiliary subunits is not understood3,7,9–12. NALCN, UNC79 and UNC80 are essential in rodents2,9,13, and mutations in human NALCN and UNC80 cause severe developmental and neurological disease14,15. Since fundamental aspects about the composition, assembly, and gating of the NALCN channelosome remain obscure, we determined the structure of this ~1 megadalton complex. UNC79 and UNC80 are massive HEAT-repeat proteins that form an intertwined anti-parallel superhelical assembly which docks intracellularly onto the NALCN-FAM155A pore-forming subcomplex. Calmodulin copurifies bound to the carboxy-terminal domain of NALCN, identifying this region as a putative modulatory hub. Single channel analyses uncovered a low open probability for the wild-type complex, highlighting the tightly closed S6-gate in the structure, and providing a basis to interpret the altered gating properties of disease-causing variants. Key constraints between the UNC79-UNC80 subcomplex and the NALCN DI-DII and DII-DIII linkers are identified that lead to a model of channelosome gating. Our results provide a structural blueprint to understand NALCN channelosome physiology and a template for drug discovery to modulate the resting membrane potential.