Peter Hegemann

Bestrhodopsins constitute a class of light-regulated pentameric ion channels that consist of one or two rhodopsins in tandem fused with bestrophin ion channel domains. Here, we report on the isomerization dynamics in the rhodopsin tandem domains of Phaeocystis antarctica bestrhodopsin, which binds all-trans retinal Schiff-base (RSB) absorbing at 661 nm and, upon illumination, converts to the meta-stable P540 state with an unusual 11- cis RSB. The primary photoproduct P682 corresponds to a mixture of highly distorted 11 -cis and 13- cis RSB directly formed from the excited state in 1.4 ps. P673 evolves from P682 in 500 ps and contains highly distorted 13- cis RSB, indicating that the 11- cis fraction in P682 converts to 13- cis . Next, P673 establishes an equilibrium with P595 in 1.2 µs, during which RSB converts to 11- cis and then further proceeds to P560 in 48 µs and P540 in 1.0 ms while remaining 11- cis . Hence, extensive isomeric switching occurs on the early ground state potential energy surface (PES) on the hundreds of ps to µs timescale before finally settling on a metastable 11- cis photoproduct. We propose that P682 and P673 are trapped high up on the ground-state PES after passing through either of two closely located conical intersections that result in 11- cis and 13- cis RSB. Co-rotation of C11=C12 and C13=C14 bonds results in a constricted conformational landscape that allows thermal switching between 11 -cis and 13- cis species of highly strained RSB chromophores. Protein relaxation may release RSB strain, allowing it to evolve to a stable 11- cis isomeric configuration in microseconds.