Photon absorption by BR initiates the isomerization of bound retinal from the all-trans to the 13-cis configuration (Fig. 2B), thereby triggering a series of proton-transfer reactions that constitute the proton translocation mechanism of BR (Fig. 2C). The conformational shift and changing dipole of the RSBH+ raises its pK, resulting in release of the RSB proton and generating the M intermediate, named after the active state of vertebrate rhodopsin, which maximally absorbs light at 410 nm; the released proton is accepted by the nearby Asp 85, which is part of the counterion complex. A distinct proton is then released from a group of amino acids near the extracellular side of the opsin, termed the proton release complex (PRC), out to the extracellular medium. At this point, the RSB captures a proton from Asp 96, and the protein enters the N intermediate, which absorbs at 560 nm. Asp 96 is then reprotonated by resident cytoplasmic protons, which is the key step in modulating cellular proton concentration. During this Asp 96 proton uptake, the Schiff base reisomerizes back to all-trans (defining the O state), which absorbs at 630 nm. In the final step, Asp 85 transfers its proton to the PRC, thereby completing one full photocycle and returning BR to its original state. The net outcome is that, upon absorption of a single photon, BR releases one proton to the extracellular milieu and absorbs a second proton from the cytoplasm in a sequence of steps that are spatially disconnected. As discussed below, amino acids involved in the proton translocation reactions are commonly conserved across other type I opsins with homology to BR and appear to function as a conserved pathway for proton translocation.
Photon absorption by BR initiates the isomerization of bound retinal from the all-trans to the 13-cis configuration (Fig. 2B), thereby triggering a series of proton-transfer reactions that constitute the proton translocation mechanism of BR (Fig. 2C). The conformational shift and changing dipole of the RSBH+ raises its pK, resulting in release of the RSB proton and generating the M intermediate, named after the active state of vertebrate rhodopsin, which maximally absorbs light at 410 nm; the released proton is accepted by the nearby Asp 85, which is part of the counterion complex. A distinct proton is then released from a group of amino acids near the extracellular side of the opsin, termed the proton release complex (PRC), out to the extracellular medium. At this point, the RSB captures a proton from Asp 96, and the protein enters the N intermediate, which absorbs at 560 nm. Asp 96 is then reprotonated by resident cytoplasmic protons, which is the key step in modulating cellular proton concentration. During this Asp 96 proton uptake, the Schiff base reisomerizes back to all-trans (defining the O state), which absorbs at 630 nm. In the final step, Asp 85 transfers its proton to the PRC, thereby completing one full photocycle and returning BR to its original state. The net outcome is that, upon absorption of a single photon, BR releases one proton to the extracellular milieu and absorbs a second proton from the cytoplasm in a sequence of steps that are spatially disconnected. As discussed below, amino acids involved in the proton translocation reactions are commonly conserved across other type I opsins with homology to BR and appear to function as a conserved pathway for proton translocation.
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