Organometallic and radical intermediates reveal mechanism of diphthamide biosynthesis.

Publication Type:

Journal Article

Source:

Science, Volume 359, Issue 6381, p.1247-1250 (2018)

Abstract:

<p>Diphthamide biosynthesis involves a carbon-carbon bond-forming reaction catalyzed by a radical S-adenosylmethionine (SAM) enzyme that cleaves a carbon-sulfur (C-S) bond in SAM to generate a 3-amino-3-carboxypropyl (ACP) radical. Using rapid freezing, we have captured an organometallic intermediate with an iron-carbon (Fe-C) bond between ACP and the enzyme&#39;s [4Fe-4S] cluster. In the presence of the substrate protein, elongation factor 2, this intermediate converts to an organic radical, formed by addition of the ACP radical to a histidine side chain. Crystal structures of archaeal diphthamide biosynthetic radical SAM enzymes reveal that the carbon of the SAM C-S bond being cleaved is positioned near the unique cluster Fe, able to react with the cluster. Our results explain how selective C-S bond cleavage is achieved in this radical SAM enzyme.</p>

PDB: 
6BXK, 6BXL, 6BXM, 6BXN, 6BXO
Detector: 
Q315
PILATUS
Beamline: 
24-ID-C
24-ID-E