Real time dynamics of gating-related conformational changes in CorA

CorA, a divalent-selective channel in the metal ion transport superfamily, is the major Mg²⁺-influx pathway in prokaryotes. CorA structures in closed (Mg²⁺-bound), and open (Mg²⁺-free) states, together with functional data showed that Mg²⁺-influx inhibits further Mg²⁺-uptake completing a regulatory feedback loop. While the closed state structure is a symmetric pentamer, the open state displayed unexpected asymmetric architectures. Using high-speed atomic force microscopy (HS-AFM), we explored the Mg²⁺-dependent gating transition of single CorA channels: HS-AFM movies during Mg²⁺-depletion experiments revealed the channel's transition from a stable Mg²⁺-bound state over a highly mobile and dynamic state with fluctuating subunits to asymmetric structures with varying degree of protrusion heights from the membrane. Our data shows that at Mg²⁺-concentration below Kd, CorA adopts a dynamic (putatively open) state of multiple conformations that imply structural rearrangements through hinge-bending in TM1. We discuss how these structural dynamics define the functional behavior of this ligand dependent channel.