title: Nanodomains of cytochrome b6f and photosystem II complexes in spinach grana thylakoid membranes.
abstract: The cytochrome b6f (cytb6f) complex plays a central role in photosynthesis, coupling electron transport between photosystem II (PSII) and photosystem I to the generation of a transmembrane proton gradient used for the biosynthesis of ATP. Photosynthesis relies on rapid shuttling of electrons by plastoquinone (PQ) molecules between PSII and cytb6f complexes in the lipid phase of the thylakoid membrane. Thus, the relative membrane location of these complexes is crucial, yet remains unknown. Here, we exploit the selective binding of the electron transfer protein plastocyanin (Pc) to the lumenal membrane surface of the cytb6f complex using a Pc-functionalized atomic force microscope (AFM) probe to identify the position of cytb6f complexes in grana thylakoid membranes from spinach (Spinacia oleracea). This affinity-mapping AFM method directly correlates membrane surface topography with Pc-cytb6f interactions, allowing us to construct a map of the grana thylakoid membrane that reveals nanodomains of colocalized PSII and cytb6f complexes. We suggest that the close proximity between PSII and cytb6f complexes integrates solar energy conversion and electron transfer by fostering short-range diffusion of PQ in the protein-crowded thylakoid membrane, thereby optimizing photosynthetic efficiency.
authors.affiliation: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, United Kingdom matt.johnson@sheffield.ac.uk.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Johnson
authors.affiliation: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, United Kingdom.
authors.forename: Cvetelin
authors.initials: C
authors.lastname: Vasilev
authors.affiliation: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, United Kingdom.
authors.forename: John D
authors.initials: JD
authors.lastname: Olsen
authors.affiliation: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, United Kingdom.
authors.forename: C Neil
authors.initials: CN
authors.lastname: Hunter
mesh_terms: D045346:Cytochrome b6f Complex / Q000378:metabolism*; D004058:Diffusion; D004579:Electron Transport; D018625:Microscopy, Atomic Force; D010084:Oxidation-Reduction; D010788:Photosynthesis; D045331:Photosystem I Protein Complex / Q000378:metabolism*; D045332:Photosystem II Protein Complex / Q000378:metabolism*; D010970:Plastocyanin / Q000378:metabolism; D010971:Plastoquinone / Q000378:metabolism; D018724:Spinacia oleracea / Q000235:genetics / Q000378:metabolism*; D020524:Thylakoids / Q000378:metabolism
chemical_list: D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D010970:Plastocyanin; D045346:Cytochrome b6f Complex; D010971:Plastoquinone