Organometallic and radical intermediates reveal mechanism of diphthamide biosynthesis

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’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.

Source: ScienceNOW - March 15, 2018 Category: Science Authors: Dong, M., Kathiresan, V., Fenwick, M. K., Torelli, A. T., Zhang, Y., Caranto, J. D., Dzikovski, B., Sharma, A., Lancaster, K. M., Freed, J. H., Ealick, S. E., Hoffman, B. M., Lin, H. Tags: Biochemistry reports Source Type: news