Biohybrid photosynthetic charge accumulation detected by flavin semiquinone formation in ferredoxin-NADP + reductase

Utschig, Lisa M.; Brahmachari, Udita; Mulfort, Karen L.; Niklas, Jens; Poluektov, Oleg G.

doi:10.1039/D2SC01546C

Title: Biohybrid photosynthetic charge accumulation detected by flavin semiquinone formation in ferredoxin-NADP ⁺ reductase

Journal Article · Tue Jun 07 00:00:00 EDT 2022 · Chemical Science

DOI:https://doi.org/10.1039/D2SC01546C· OSTI ID:1867522

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Chemical Sciences and Engineering Division, Argonne National Laboratory, Lemont, IL 60439, USA

Flavin chemistry is ubiquitous in biological systems with flavoproteins engaged in important redox reactions. In photosynthesis, flavin cofactors are used as electron donors/acceptors to facilitate charge transfer and accumulation for ultimate use in carbon fixation. Following light-induced charge separation in the photosynthetic transmembrane reaction center photosystem I (PSI), an electron is transferred to one of two small soluble shuttle proteins, a ferredoxin (Fd) or a flavodoxin (Fld) (the latter in the condition of Fe-deficiency), followed by electron transfer to the ferredoxin-NADP+ reductase (FNR) enzyme. FNR accepts two of these sequential one electron transfers, with its flavin adenine dinucleotide (FAD) cofactor becoming doubly reduced, forming a hydride which is then passed onto the substrate NADP⁺ to form NADPH. The two one-electron potentials (oxidized/semiquinone and semiquinone/hydroquinone) are similar to each other with the FNR protein stabilizing the hydroquinone, making spectroscopic detection of the intermediate semiquinone state difficult. We employed a new biohybrid-based strategy that involved truncating the native three-protein electron transfer cascade PSI → Fd → FNR to a two-protein cascade by replacing PSI with a molecular Ru(II) photosensitizer (RuPS) which is covalently bound to Fd and Fld to form biohybrid complexes that successfully mimic PSI in light-driven NADPH formation. RuFd → FNR and RuFld → FNR electron transfer experiments revealed a notable distinction in photosynthetic charge accumulation that we attribute to the different protein cofactors [2Fe2S] and flavin. After freeze quenching the two-protein systems under illumination, an intermediate semiquinone state of FNR was readily observed with cw X-band EPR spectroscopy. The increased spectral resolution from selective deuteration allowed EPR detection of inter-flavoprotein electron transfer. This work establishes a biohybrid experimental approach for further studies of photosynthetic light-driven electron transfer chain that culminates at FNR and highlights nature's mechanisms that couple single electron transfer chemistry to charge accumulation, providing important insight for the development of photon-to-fuel schemes.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences & Biosciences Division

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1867522

Alternate ID(s):: OSTI ID: 1879891

Journal Information:: Chemical Science, Journal Name: Chemical Science Vol. 13 Journal Issue: 22; ISSN 2041-6520

Publisher:: Royal Society of Chemistry (RSC)Copyright Statement

Country of Publication:: United Kingdom

Language:: English

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