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The radiation-resistant bacterium Deinococcus radiodurans is known as the world’s toughest bacterium. The S-layer of D. radiodurans, consisting of several proteins on the surface of the cellular envelope and intimately associated with the outer membrane, has therefore been useful as a model for structural and functional studies. Its main proteinaceous unit, the S-layer deinoxanthin-binding complex (SDBC), is a hetero-oligomeric assembly known to contribute to the resistance against environmental stress and have porin functional features; however, its precise structure is unknown. Here, we resolved the structure of the SDBC at (2.5 Å resolution by cryo-EM and assigned the sequence of its main subunit, the protein DR_2577. This structure is characterized by a pore region, a massive β-barrel organization, a stalk region consisting of a trimeric coiled coil, and a collar region at the base of the stalk. We show that each monomer binds three Cu ions and one Fe ion and retains one deinoxanthin molecule and two phosphoglycolipids, all exclusive to D. radiodurans. Finally, electrophysiological characterization of the SDBC shows that it exhibits transport properties with several amino acids. Taken together, these results highlight the SDBC as a robust structure displaying both protection and sieving functions that facilitates exchanges with the environment. [ABSTRACT FROM AUTHOR]

We report the fabrication of an oriented bioelectrode of photosystem I (PSI) on single-layer graphene (SLG). This bioelectrode demonstrates improved photocurrent generation, which can be directly attributed to the molecular conductive interface formed by cytochrome c553 (cyt c553) promoting the uniform orientation of PSI with its donor side towards the electrode. The conductive interface between PSI-cyt c553 and SLG is facilitated by a monolayer composed of π-π-stacked pyrene functionalized with the Ni-NTA moiety, which binds the His6-tagged cyt c553. The surface uniformity of the PSI protein orientation in the electrode structure is evidenced by cross-sectional scanning electron microscopy and fluorescence microscopy, with the latter also proving the efficient electronic coupling between majority of the PSI complexes and graphene. With the uniform organization of the biological photoactive layer, photocurrents are generated at the open circuit potential, which can be further increased when a negative potential is applied. Indeed, at the highest applied negative potential (-0.3 V), over 5-fold increase in the cathodic photocurrent for the PSI complexes conjugated via cyt c553 to the SLG substrate is observed compared with that obtained for the randomly oriented structure where PSI is physisorbed on graphene. These results indicate the key role of a strictly defined orientation of photoactive proteins on electrodes for proper electron transfer and substantial improvement in photocurrent generation in the present or similar bioelectrode architectures. [ABSTRACT FROM AUTHOR]

The monomeric photosystem I-light-harvesting antenna complex I (PSI-LHCI) supercomplex from the extremophilic red alga Cyanidioschyzon merolae represents an intermediate evolutionary link between the cyanobacterial PSI reaction center and its green algal/higher plant counterpart. We show that the C. merolae PSI-LHCI supercomplex is characterized by robustness in various extreme conditions. By a combination of biochemical, spectroscopic, mass spectrometry, and electron microscopy/single particle analyses, we dissected three molecular mechanisms underlying the inherent robustness of the C. merolae PSI-LHCI supercomplex: (1) the accumulation of photoprotective zeaxanthin in the LHCI antenna and the PSI reaction center; (2) structural remodeling of the LHCI antenna and adjustment of the effective absorption cross section; and (3) dynamic readjustment of the stoichiometry of the two PSI-LHCI isomers and changes in the oligomeric state of the PSI-LHCI supercomplex, accompanied by dissociation of the PsaK core subunit. We show that the largest low light-treated C. merolae PSI-LHCI supercomplex can bind up to eight Lhcr antenna subunits, which are organized as two rows on the PsaF/PsaJ side of the core complex. Under our experimental conditions, we found no evidence of functional coupling of the phycobilisomes with the PSI-LHCI supercomplex purified from various light conditions, suggesting that the putative association of this antenna with the PSI supercomplex is absent or may be lost during the purification procedure. [ABSTRACT FROM AUTHOR]