Thylakoid membranes in the chloroplast of plants, algae, and cyanobacteria are the powerhouse of the photosynthesis capturing the solar energy and converting it into chemical energies. Although their overall structures and functions have been extensively studied, the intrinsically heterogeneous and dynamic two-dimensional (2D) membrane structures are not fully understood. Investigating native thylakoid membranes in vivo is difficult due to their small sizes and limited external access to the chloroplast interior, while the bottom-up approaches based on model systems have been hampered by the sheer complexity of the native membrane. Here we try to fill the gap by reconstituting the whole thylakoid membrane into a patterned substrate-supported planer bilayer. A mixture of thylakoid membrane purified from spinach and synthetic phospholipid (DOPC) vesicles spontaneously formed a laterally continuous and fluid 2D membrane in the scaffold of patterned polymeric bilayer. Chlorophyll fluorescence arising from photosystem II (PSII) recovered after photobleaching, suggesting that the membrane components are laterally mobile. The electron transfer activity of PSII was retained as confirmed by the changes in chlorophyll fluorescence in the presence of the electron acceptors and/ or inhibitors. Furthermore, we confirmed the electron transfer activity of photosystem I (PSI) by observing the generation of NADPH in the presence of water-soluble ferredoxin and ferredoxin-NADP<sup>+</sup> reductase. The lateral mobility of membrane-bound molecules and the functional reconstitution of major photosystems support the feasibility of reconstituted thylakoid-synthetic lipid hybrid membranes as the experimental platform to study the 2D organization and molecular machinery of photosynthesis.

Hybrid photosynthetic system of natural plant thylakoids and synthetic lipids reconstituted into a supported microscale membrane array

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.