The coupled photocycle of phenyl-p-benzoquinone and Light-Harvesting Complex II (LHCII) within the biohybrid system.

The combination of trimeric form of the light-harvesting complex II (LHCII3), a porous graphite electrode (GE), and the application of phenyl-p-benzoquinone (PPBQ), the quinone derivative, allow the construction of a new type of biohybrid photoactive system. The Chl fluorescence decay and voltammetric analyzes revealed that PPBQ impacts LHCII3 proportionally to accessible quenching sites and that PPBQ forms redox complexes with Chl in both ground and excited states. As a result, photocurrent generation is directly dependent on PPBQ-induced quenching of Chl fluorescence. Since PPBQ also undergoes photoactivation, the action of GE-LHCII3-PPBQ depends on the mutual coupling of LHCII3 and PPBQ photocycles. The GE-LHCII3-PPBQ generates a photocurrent of up to 4.5 µA and exhibits considerable stability during operation. The three-dimensional arrangement of graphite scraps in GE builds an active electrode surface and stabilizes LHCII3 in its native form in low-density multilayers. The results indicate the future usability of such designed photoactive device.

Comparison of α-Helix and ß-Sheet Structure Adaptation to a Quantum Dot Geometry: Toward the Identification of an Optimal Motif for a Protein Nanoparticle Cover.

While quantum dots (QDs) are useful as fluorescent labels, their application in biosciences is limited due to the stability and hydrophobicity of their surface. In this study, we tested two types of proteins for use as a cover for spherical QDs, composed of cadmium selenide. Pumilio homology domain (Puf), which is mostly α-helical, and leucine-rich repeat (LRR) domain, which is rich in ß-sheets, were selected to determine if there is a preference for one of these secondary structure types for nanoparticle covers. The protein sequences were optimized to improve their interaction with the surface of QDs. The solubilization of the apoproteins and their assembly with nanoparticles required the application of a detergent, which was removed in subsequent steps. Finally, only the Puf-based cover was successful enough as a QD hydrophilic cover. We showed that a single polypeptide dimer of Puf, PufPuf, can form a cover. We characterized the size and fluorescent properties of the obtained QD:protein assemblies. We showed that the secondary structure of the Puf proteins was not destroyed upon contact with the QDs. We demonstrated that these assemblies do not promote the formation of reactive oxygen species during illumination of the nanoparticles. The data represent advances in the effort to obtain a stable biocompatible cover for QDs.

Electrochemical characterization of LHCII on graphite electrodes - Potential-dependent photoactivation and arrangement of complexes.

Light-dependent electrochemical properties of the light harvesting complexes of Photosystem II (LHCII) and the corresponding interactions with screen-printed graphite electrodes (GEs) are determined. No exogenous soluble redox mediators are used. LHCII isolated from spinach leaves are immobilized on GE by physical adsorption and through interactions with glutaraldehyde. Importantly, the insertion of LHCII into the pores of a GE is achieved by subjecting the electrode to specific potentials. Both trimeric and aggregated forms of LHCII located within the graphite layer retain their native structures. Voltammetric current peaks centred at ca. -230 and + 50 mV vs. Ag/AgCl (+94 and + 374 mV vs. NHE) limit the investigation of the reduction and oxidation processes of immobilized LHCII. An anodic photocurrent is generated in the LHCII-GE proportional to light intensity and can reach a value of 150 nA/cm2. Light-dependent charge separation in LHCII followed by electron transfer to the GE occurs only at potentials of above -200 mV vs. Ag/AgCl (+124 mV vs. NHE). Our results illustrate the importance of the structural proximity of LHCII and GE for photocurrent generation.