Intramolecular seal against proton backflow in bacteriorhodopsin

Many microorganisms can harvest energy from sunlight to establish an electrochemical potential gradient across cell membranes by pumping protons. In addition to their pumping mechanisms, these ion pumps must also effectively counter the constant threat of gradient-driven backflow at the molecular level. A recent study (Ren, PNAS Nexus 1, pgac103, 2022) unscrambled the mixed structural signals in femtosecond serial crystallographic datasets and delineated the molecular mechanism of retinal photoisomerization in bacteriorhodopsin, a model system for light-driven proton pumps. This study continues to resolve structural heterogeneity, substantially improves the quality of electron density maps, and provides a clear visualization of the isolated intermediate states from I to M. These intermediate structures reveal the motion of an intramolecular seal at work, which constantly partitions the cytoplasmic and extracellular half-channels, thus preventing proton backflow.