7ndw
ThyX-FADH2 soaked with 20 mM FormaldehydeThyX-FADH2 soaked with 20 mM Formaldehyde
Structural highlights
FunctionTHYX_THEMA Catalyzes the formation of dTMP and tetrahydrofolate from dUMP and methylenetetrahydrofolate. Publication Abstract from PubMedFolate enzyme cofactors and their derivatives have the unique ability to provide a single carbon unit at different oxidation levels for the de novo synthesis of amino-acids, purines, or thymidylate, an essential DNA nucleotide. How these cofactors mediate methylene transfer is not fully settled yet, particularly with regard to how the methylene is transferred to the methylene acceptor. Here, we uncovered that the bacterial thymidylate synthase ThyX, which relies on both folate and flavin for activity, can also use a formaldehyde-shunt to directly synthesize thymidylate. Combining biochemical, spectroscopic and anaerobic crystallographic analyses, we showed that formaldehyde reacts with the reduced flavin coenzyme to form a carbinolamine intermediate used by ThyX for dUMP methylation. The crystallographic structure of this intermediate reveals how ThyX activates formaldehyde and uses it, with the assistance of active site residues, to methylate dUMP. Our results reveal that carbinolamine species promote methylene transfer and suggest that the use of a CH2O-shunt may be relevant in several other important folate-dependent reactions. An enzymatic activation of formaldehyde for nucleotide methylation.,Bou-Nader C, Stull FW, Pecqueur L, Simon P, Guerineau V, Royant A, Fontecave M, Lombard M, Palfey BA, Hamdane D Nat Commun. 2021 Jul 27;12(1):4542. doi: 10.1038/s41467-021-24756-8. PMID:34315871[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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