[FeFe]-Hydrogenase: Defined Lysate-Free Maturation Reveals a Key Role for Lipoyl-H-Protein in DTMA Ligand Biosynthesis

Pagnier, Adrien; Balci, Batuhan; Shepard, Eric M.; Yang, Hao; Warui, Douglas M.; Impano, Stella; Booker, Squire J.; Hoffman, Brian M.; Broderick, William E.; Broderick, Joan B.

doi:10.1002/anie.202203413

Title: [FeFe]-Hydrogenase: Defined Lysate-Free Maturation Reveals a Key Role for Lipoyl-H-Protein in DTMA Ligand Biosynthesis

Journal Article · 23 March 2022 · Angewandte Chemie (International Edition)

DOI: https://doi.org/10.1002/anie.202203413 · OSTI ID:1976282

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Montana State University, Bozeman, MT (United States)
Northwestern University, Evanston, IL (United States)
Pennsylvania State University, University Park, PA (United States)
Pennsylvania State University, University Park, PA (United States); Howard Hughes Medical Institute, Chevy Chase, MD (United States)
Department of Chemistry & Biochemistry Montana State University Bozeman MT 59717 USA

Abstract Maturation of [FeFe]‐hydrogenase (HydA) involves synthesis of a CO, CN ⁻ , and dithiomethylamine (DTMA)‐coordinated 2Fe subcluster that is inserted into HydA to make the active hydrogenase. This process requires three maturation enzymes: the radical S‐adenosyl‐ l ‐methionine (SAM) enzymes HydE and HydG, and the GTPase HydF. In vitro maturation with purified maturation enzymes has been possible only when clarified cell lysate was added, with the lysate presumably providing essential components for DTMA synthesis and delivery. Here we report maturation of [FeFe]‐hydrogenase using a fully defined system that includes components of the glycine cleavage system (GCS), but no cell lysate. Our results reveal for the first time an essential role for the aminomethyl‐lipoyl‐H‐protein of the GCS in hydrogenase maturation and the synthesis of the DTMA ligand of the H‐cluster. In addition, we show that ammonia is the source of the bridgehead nitrogen of DTMA.

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Research Organization:: Montana State Univ., Bozeman, MT (United States)

Sponsoring Organization:: USDOE Office of Science (SC); National Science Foundation (NSF); National Institutes of Health (NIH); MJ MurdockCharitable Trust; USDOE

Grant/Contract Number:: SC0005404; MCB-1716686; GM-111097; P20GM103474; S10OD28650

OSTI ID:: 1976282

Alternate ID(s):: OSTI ID: 1862709

Journal Information:: Angewandte Chemie (International Edition), Vol. 61, Issue 22; ISSN 1433-7851

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
hydrogenase maturation
lipoyl-H-protein
biosynthesis
glycine cleavage system
dithiomethylamine

Title: [FeFe]-Hydrogenase: Defined Lysate-Free Maturation Reveals a Key Role for Lipoyl-H-Protein in DTMA Ligand Biosynthesis

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