Archaeal and eukaryotic translation elongation factor 2 contain a unique post-translationally modified histidine residue called diphthamide, which is the target of diphtheria toxin. The biosynthesis of diphthamide was proposed to involve three steps, with the first being the formation of a C–C bond between the histidine residue and the 3-amino-3-carboxypropyl group of S-adenosyl-l-methionine (SAM). However, further details of the biosynthesis remain unknown. Here we present structural and biochemical evidence showing that the first step of diphthamide biosynthesis in the archaeon Pyrococcus horikoshii uses a novel iron–sulphur-cluster enzyme, Dph2. Dph2 is a homodimer and each of its monomers can bind a [4Fe–4S] cluster. Biochemical data suggest that unlike the enzymes in the radical SAM superfamily, Dph2 does not form the canonical 5′-deoxyadenosyl radical. Instead, it breaks the Cγ,Met–S bond of SAM and generates a 3-amino-3-carboxypropyl radical. Our results suggest that P. horikoshii Dph2 represents a previously unknown, SAM-dependent, [4Fe–4S]-containing enzyme that catalyses unprecedented chemistry.
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We thank L. Kinsland for assistance with manuscript preparation, the Dreyfus Foundation for a New Faculty Award to H.L., NIH/NIGMS R01GM088276 to H.L. and S.E.E., and NIH/NCRR P41-RR016292 for the ACERT Center Grant to J.F. This work is based upon research conducted at the Advanced Photon Source (APS), Argonne National Laboratory, on the Northeastern Collaborative Access Team beamlines, which are supported by award RR-15301 from the US National Center for Research Resources at the US National Institutes of Health. Use of the APS is supported by the US Department of Energy, Office of Basic Energy Sciences, under contract no. DE-AC02-06CH11357.
The authors declare no competing financial interests.
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Zhang, Y., Zhu, X., Torelli, A. et al. Diphthamide biosynthesis requires an organic radical generated by an iron–sulphur enzyme. Nature 465, 891–896 (2010). https://doi.org/10.1038/nature09138
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