Download MendeleyRevealing reaction intermediates in one-carbon elongation by thiamine diphosphate/CoA-dependent enzyme family.
Kim, Y., Lee, S.H., Gade, P., Nattermann, M., Maltseva, N., Endres, M., Chen, J., Wichmann, P., Hu, Y., Marchal, D.G., Yoshikuni, Y., Erb, T.J., Gonzalez, R., Michalska, K., Joachimiak, A.(2024) Commun Chem 7: 160-160
- PubMed: 39034323
- DOI: https://doi.org/10.1038/s42004-024-01242-y
- Primary Citation of Related Structures:
8VZA, 8VZB, 8VZC, 8VZD, 8VZE, 8VZF, 8VZH, 8VZI, 8VZJ, 8VZK - PubMed Abstract:
2-Hydroxyacyl-CoA lyase/synthase (HACL/S) is a thiamine diphosphate (ThDP)-dependent versatile enzyme originally discovered in the mammalian ¦Á-oxidation pathway. HACL/S natively cleaves 2-hydroxyacyl-CoAs and, in its reverse direction, condenses formyl-CoA with aldehydes or ketones. The one-carbon elongation biochemistry based on HACL/S has enabled the use of molecules derived from greenhouse gases as biomanufacturing feedstocks. We investigated several HACL/S family members with high activity in the condensation of formyl-CoA and aldehydes, and distinct chain-length specificities and kinetic parameters. Our analysis revealed the structures of enzymes in complex with acyl-CoA substrates and products, several covalent intermediates, bound ThDP and ADP, as well as the C-terminal active site region. One of these observed states corresponds to the intermediary ¦Á-carbanion with hydroxymethyl-CoA covalently attached to ThDP. This research distinguishes HACL/S from related sub-families and identifies key residues involved in substrate binding and catalysis. These findings expand our knowledge of acyloin-condensation biochemistry and offer attractive prospects for biocatalysis using carbon elongation.
Organizational Affiliation:
eBERlight and Structural Biology Center, X-ray Science Division, Argonne National Laboratory, Lemont, IL, USA.
