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Low-barrier hydrogen bond powers long-range radical transfer in the metal-free ribonucleotide reductase
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics. Indian Institute of Science, India.ORCID iD: 0000-0002-4073-7627
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics. Technical University Dortmund, Germany.ORCID iD: 0000-0001-9626-3670
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics.ORCID iD: 0000-0001-5033-2810
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Number of Authors: 132026 (English)In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 123, no 19, article id e2529856123Article in journal (Refereed) Published
Abstract [en]

Ribonucleotide reductases (RNRs) catalyze the conversion of ribonucleotide (RNA) to deoxyribonucleotide (DNA) building blocks initiated by a long-range (>30 Å) proton-coupled electron transfer (PCET) by mechanistic principles that remain much debated. By combining multiscale quantum and classical simulations with directed mutagenesis, X-ray crystallography, and vibrational and electron paramagnetic resonance spectroscopy, we elucidate here the molecular principles underlying how metal-free RNRs initiate the long-range PCET process by creating a highly stable 3,4-dihydroxyphenylalanine (DOPA) initiator radical. We show that DOPA• is redox-tuned by a low-barrier hydrogen bond (LBHB), with a delocalized proton that provides the catalytic power for the ribonucleotide reduction. We find that the LBHB couples to an extended hydrogen-bonded network, with distant mutations resulting in the loss of radical formation, and providing key molecular insight into the long-range radical transport mechanism in RNRs. On a general level, our findings support the direct involvement of LBHB in protein chemistry and the importance of quantum effects in enzyme catalysis.

Place, publisher, year, edition, pages
2026. Vol. 123, no 19, article id e2529856123
Keywords [en]
LBHB, PCET, QM/MM XFEL, quantum biology
National Category
Molecular Biology
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URN: urn:nbn:se:su:diva-256172DOI: 10.1073/pnas.2529856123PubMedID: 42096306Scopus ID: 2-s2.0-105038372933OAI: oai:DiVA.org:su-256172DiVA, id: diva2:2065921
Available from: 2026-06-04 Created: 2026-06-04 Last updated: 2026-06-04Bibliographically approved

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Sirohiwal, AbhishekJohn, JulianeKumar, RohitSrinivas, VivekPöverlein, Maximilian C.Gamiz-Hernandez, Ana P.Högbom, MartinKaila, Ville R. I.

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Sirohiwal, AbhishekJohn, JulianeKumar, RohitSrinivas, VivekPöverlein, Maximilian C.Gamiz-Hernandez, Ana P.Högbom, MartinKaila, Ville R. I.
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Department of Biochemistry and Biophysics
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Proceedings of the National Academy of Sciences of the United States of America
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