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.
Original language | English (US) |
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Article number | e202203413 |
Journal | Angewandte Chemie - International Edition |
Volume | 61 |
Issue number | 22 |
DOIs | |
State | Published - May 23 2022 |
Funding
This work was funded by the U.S. DOE (DE\u2010SC0005404 to J.B.B. and E.M.S.). S.J.B acknowledges funding from the NSF (MCB\u20101716686), and B.M.H from the NIH GM 111097. The Mass Spectrometry Facility was in part funded by the MJ Murdock Charitable Trust and the NIH (P20GM103474 and S10OD28650). The authors thank Prof. Inger Andersson (Uppsala U.) for the gift of pBAD\u2010HisA\u2010slr0293, and Juan Fontecilla\u2010Camps, Roman Rohac, and Yvain Nicolet for helpful discussions.
Keywords
- Biosynthesis
- Dithiomethylamine
- Glycine Cleavage System
- Hydrogenase Maturation
- Lipoyl-H-Protein
ASJC Scopus subject areas
- Catalysis
- General Chemistry