D-Band EPR and ENDOR Spectroscopy of 15N-Labeled Photosystem I

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

doi:10.1007/s00723-021-01438-8

Title: D-Band EPR and ENDOR Spectroscopy of ¹⁵N-Labeled Photosystem I

Journal Article · Sat Nov 20 00:00:00 EST 2021 · Applied Magnetic Resonance

DOI:https://doi.org/10.1007/s00723-021-01438-8· OSTI ID:1902996

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Argonne National Lab. (ANL), Argonne, IL (United States)

For billions of years, nature has optimized the photosynthetic machinery that converts light energy into chemical energy. Key primary reactions of photosynthesis occur in large membrane protein-cofactor complexes. The light-induced sequential electron transfer reactions occur through a chain of donor/acceptor cofactors embedded in the protein matrix resulting in a long-lived transmembrane charge-separated state. EPR is the method of choice to study electron transfer and the interaction of protein environment with redox-active cofactors. However, the spectra of organic cofactor radicals typically are not fully resolved and severely overlap at conventional X-band EPR. Even at Q-band EPR, this overlap is present and often a serious problem. As a result, there is a large variation of the reported EPR data and limited understanding of electronic structures of several redox-active cofactors. These serious problems can often be overcome by the excellent spectral resolution provided by high-frequency EPR (HF EPR). In this work, we study the electronic structure of the primary electron donor P₇₀₀ and the secondary electron acceptor A₁ of Photosystem I (PSI) using 130 GHz (D-band) EPR and Electron-Nuclear-Double-Resonance (ENDOR) spectroscopy. PSI was isotopically labeled with ¹⁵N (I = $$\frac{1/2}$$) to avoid quadrupolar interactions in the most abundant nitrogen isotope ¹⁴N (I = 1) and simplify the ENDOR spectra. ENDOR spectroscopy is central for determining the hyperfine coupling of nitrogen atoms of the two chlorophyll molecules comprising oxidized P₇₀₀ and the involvement of protein nitrogen atoms with reduced A₁. While HF ENDOR of A₁^- allows identification of two nitrogen atoms, HF ENDOR of P₇₀₀⁺ still does not permit unique assignment of the recorded hyperfine couplings.

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

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

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1902996

Journal Information:: Applied Magnetic Resonance, Vol. 53, Issue 7-9; ISSN 0937-9347

Publisher:: Springer NatureCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Photosynthesis
Photosystem I
High-frequency EPR
ENDOR
isotope labeling
15N

Title: D-Band EPR and ENDOR Spectroscopy of 15N-Labeled Photosystem I

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Title: D-Band EPR and ENDOR Spectroscopy of ¹⁵N-Labeled Photosystem I