Publication Date:
2016-01-30
Description:
In several human pathogens, thyX-encoded flavin-dependent thymidylate synthase (FDTS) catalyzes the last step in the biosynthesis of thymidylate, one of the four DNA nucleotides. ThyX is absent in humans, rendering FDTS an attractive antibiotic target; however, the lack of mechanistic understanding prohibits mechanism-based drug design. Here, we report trapping and characterization of two consecutive intermediates, which together with previous crystal structures indicate that the enzyme's reduced flavin relays a methylene from the folate carrier to the nucleotide acceptor. Furthermore, these results corroborate an unprecedented activation of the nucleotide that involves no covalent modification but only electrostatic polarization by the enzyme's active site. These findings indicate a mechanism that is very different from thymidylate biosynthesis in humans, underscoring the promise of FDTS as an antibiotic target.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4744818/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4744818/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Mishanina, Tatiana V -- Yu, Liping -- Karunaratne, Kalani -- Mondal, Dibyendu -- Corcoran, John M -- Choi, Michael A -- Kohen, Amnon -- R01 GM110775/GM/NIGMS NIH HHS/ -- T32 GM008365/GM/NIGMS NIH HHS/ -- New York, N.Y. -- Science. 2016 Jan 29;351(6272):507-10. doi: 10.1126/science.aad0300.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Chemistry, University of Iowa, Iowa City, IA 52242, USA. amnon-kohen@uiowa.edu. ; Nuclear Magnetic Resonance (NMR) Core Facility and Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA. ; Department of Chemistry, University of Iowa, Iowa City, IA 52242, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/26823429" target="_blank"〉PubMed〈/a〉
Keywords:
Bacterial Proteins/*chemistry
;
Catalysis
;
Catalytic Domain
;
*DNA Methylation
;
Flavins/chemistry
;
Folic Acid/chemistry
;
Folic Acid Transporters/chemistry
;
Humans
;
Kinetics
;
Thermotoga maritima/enzymology
;
Thymidine Monophosphate/*biosynthesis/chemistry
;
Thymidylate Synthase/*chemistry
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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