Observation of anomalous carotenoid and blind chlorophyll activations in photosystem I under synthetic membrane confinements

The role of natural thylakoid membrane confinements in architecting the robust structural and electrochemical properties of PSI is not fully understood. Most PSI studies till date extract the proteins from their natural confinements that can lead to non-native conformations. Recently our group had successfully reconstituted PSI in synthetic lipid membranes using detergent-mediated liposome solubilizations. In this study, we investigate the alterations in chlorophylls and carotenoids interactions and reorganization in PSI based on spectral property changes induced by its confinement in anionic DPhPG and zwitterionic DPhPC phospholipid membranes. To this end, we employ a combination of absorption, fluorescence, and circular dichroism (CD) spectroscopic measurements. Our results indicate unique activation and alteration of photoresponses from the PSI carotenoid (Car) bands in PSI-DPhPG proteoliposomes that can tune the Excitation Energy Transfer (EET), otherwise absent in PSI at non-native environments. Specifically, we observe broadband light harvesting via enhanced absorption in the otherwise non-absorptive green region (500–580 nm) of the Chlorophylls (Chl) along with ~64% increase in the full-width half maximum of the Qy band (650–720 nm). The CD results indicate enhanced Chl-Chl and Chl-Car interactions along with conformational changes in protein secondary structures. Such distinct changes in the Car and Chl bands are not observed in PSI confined in DPhPC. The fundamental insights into membrane microenvironments tailoring PSI subunits reorganization and interactions provide novel strategies for tuning photoexcitation processes and rational designing of biotic-abiotic interfaces in PSI-based photoelectrochemical energy conversion systems.