Neorhodopsin (NeoR) is a newly discovered fungal bistable rhodopsin that reversibly photoswitches between UV- and near-IR absorbing states denoted NeoR367 and NeoR690, respectively. NeoR367 represents a deprotonated retinal Schiff base (RSB), while NeoR690 represents a protonated RSB. Cryo-EM studies indicate that NeoR forms homodimers with 29 Å center-to-center distance between the retinal chromophores. UV excitation of NeoR367 takes place to an optically allowed S3 state of 1Bu+ symmetry, which rapidly converts to a low-lying optically forbidden S1 state of 2Ag- symmetry in 39 fs, followed by a multiexponential decay to the ground state on the 1-100 ps time scale. A theoretically predicted nπ* (S2) state does not get populated in any appreciable transient concentration during the excited-state relaxation cascade. We observe an intradimer retinal to retinal excitation energy transfer (EET) process from the NeoR367 S1 state to NeoR690, in competition with photoproduct formation. To quantitatively assess the EET mechanism and rate, we experimentally addressed and modeled the EET process under varying NeoR367-NeoR690 photoequilibrium conditions and determined the EET rate at (200 ps)-1. The NeoR367 S1 state shows a weak stimulated emission band in the near-IR around 700 nm, which may result from mixing with an intramolecular charge-transfer (ICT) state, enhancing the transition dipole moment of the S1-S0 transition and possibly facilitating the EET process. We suggest that EET may bear general relevance to the function of bistable multiwavelength rhodopsin oligomers.

• MeSH
• Dimerization MeSH
• Models, Molecular MeSH
• Protein Multimerization MeSH
• Energy Transfer MeSH
• Retinaldehyde * chemistry   MeSH
• Rhodopsins, Microbial * chemistry   metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Retinaldehyde * MeSH
• Rhodopsins, Microbial * MeSH

Cyanobacteria use large antenna complexes called phycobilisomes (PBSs) for light harvesting. However, intense light triggers non-photochemical quenching, where the orange carotenoid protein (OCP) binds to PBS, dissipating excess energy as heat. The mechanism of efficiently transferring energy from phycocyanobilins in PBS to canthaxanthin in OCP remains insufficiently understood. Using cryo-electron microscopy, we unveiled the OCP-PBS complex structure at 1.6- to 2.1-angstrom resolution, showcasing its inherent flexibility. Using multiscale quantum chemistry, we disclosed the quenching mechanism. Identifying key protein residues, we clarified how canthaxanthin's transition dipole moment in its lowest-energy dark state becomes large enough for efficient energy transfer from phycocyanobilins. Our energy transfer model offers a detailed understanding of the atomic determinants of light harvesting regulation and antenna architecture in cyanobacteria.

• MeSH
• Bacterial Proteins metabolism   MeSH
• Cryoelectron Microscopy MeSH
• Phycobilisomes * chemistry   metabolism   MeSH
• Canthaxanthin metabolism   MeSH
• Cyanobacteria * metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Names of Substances
• Bacterial Proteins MeSH
• Phycobilisomes * MeSH
• Canthaxanthin MeSH

Photosynthetic organisms harvest light using pigment-protein complexes. In cyanobacteria, these are water-soluble antennae known as phycobilisomes (PBSs). The light absorbed by PBS is transferred to the photosystems in the thylakoid membrane to drive photosynthesis. The energy transfer between these complexes implies that protein-protein interactions allow the association of PBS with the photosystems. However, the specific proteins involved in the interaction of PBS with the photosystems are not fully characterized. Here, we show in Synechocystis sp. PCC 6803 that the recently discovered PBS linker protein ApcG (sll1873) interacts specifically with PSII through its N-terminal region. Growth of cyanobacteria is impaired in apcG deletion strains under light-limiting conditions. Furthermore, complementation of these strains using a phospho-mimicking version of ApcG causes reduced growth under normal growth conditions. Interestingly, the interaction of ApcG with PSII is affected when a phospho-mimicking version of ApcG is used, targeting the positively charged residues interacting with the thylakoid membrane, suggesting a regulatory role mediated by phosphorylation of ApcG. Low-temperature fluorescence measurements showed decreased PSI fluorescence in apcG deletion and complementation strains. The PSI fluorescence was the lowest in the phospho-mimicking complementation strain, while the pull-down experiment showed no interaction of ApcG with PSI under any tested condition. Our results highlight the importance of ApcG for selectively directing energy harvested by the PBS and imply that the phosphorylation status of ApcG plays a role in regulating energy transfer from PSII to PSI.

• MeSH
• Photosystem I Protein Complex metabolism   MeSH
• Photosystem II Protein Complex metabolism   MeSH
• Phycobilisomes metabolism   MeSH
• Energy Transfer physiology   MeSH
• Synechocystis * metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Photosystem I Protein Complex MeSH
• Photosystem II Protein Complex MeSH
• Phycobilisomes MeSH