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UniProt functional annotation for P9WG57 | ||
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| UniProt code: P9WG57. |
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| Organism: | |
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv). |
| Taxonomy: | |
Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae; Mycobacterium; Mycobacterium tuberculosis complex. |
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| Function: | |
Catalyzes the reductive methylation of 2'-deoxyuridine-5'- monophosphate (dUMP) to 2'-deoxythymidine-5'-monophosphate (dTMP) while utilizing 5,10-methylenetetrahydrofolate (mTHF) as the methyl donor, and NADPH and FADH(2) as the reductant (PubMed:18493582). Is essential for growth of the pathogen on solid media in vitro; the essential function is something other than dTMP synthase (PubMed:12657046) (PubMed:22034487). {ECO:0000269|PubMed:12657046, ECO:0000269|PubMed:16139296, ECO:0000269|PubMed:18493582, ECO:0000269|PubMed:22034487}. |
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| Catalytic activity: | |
Reaction=(6R)-5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP + H(+) + NADPH = (6S)-5,6,7,8-tetrahydrofolate + dTMP + NADP(+); Xref=Rhea:RHEA:29043, ChEBI:CHEBI:15378, ChEBI:CHEBI:15636, ChEBI:CHEBI:57453, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:63528, ChEBI:CHEBI:246422; EC=2.1.1.148; Evidence={ECO:0000269|PubMed:18493582}; |
| Cofactor: | |
Name=FAD; Xref=ChEBI:CHEBI:57692; Evidence={ECO:0000269|PubMed:16139296}; Note=Binds 4 FAD per tetramer. Each FAD binding site is formed by three monomers. {ECO:0000269|PubMed:16139296}; |
| Activity regulation: | |
Is potently inhibited by 5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP), but in contrast to ThyA, is not inhibited by the folate-based 1843U89 (PubMed:18493582). A 5-alkynyl dUMP analog has been shown to highly inhibit ThyX (IC(50) value of 0.91 uM), while lacking activity against the classical mycobacterial thymidylate synthase ThyA, and therefore is a selective mycobacterial FDTS inhibitor (PubMed:21657202). {ECO:0000269|PubMed:18493582, ECO:0000269|PubMed:21657202}. |
| Biophysicochemical properties: | |
Kinetic parameters: KM=3 uM for dUMP {ECO:0000269|PubMed:18493582}; KM=4 uM for 5,10-methylenetetrahydrofolate {ECO:0000269|PubMed:18493582}; KM=47 uM for NADPH {ECO:0000269|PubMed:18493582}; Note=kcat is 0.4 min(-1). {ECO:0000269|PubMed:18493582}; |
| Pathway: | |
Pyrimidine metabolism; dTTP biosynthesis. {ECO:0000255|HAMAP- Rule:MF_01408}. |
| Subunit: | |
Homotetramer. {ECO:0000269|PubMed:16139296, ECO:0000269|PubMed:18192395, ECO:0000269|PubMed:18493582}. |
| Induction: | |
Is expressed under the exponential phase of growth, and down-regulated upon starvation. Expression of thyX is significantly increased within murine macrophages or under acid stress. Is expressed at a lower level than thyA under all of the in vitro and in vivo growth conditions tested. {ECO:0000269|PubMed:22034487}. |
| Disruption phenotype: | |
Cells lacking this gene display impaired growth (PubMed:12657046). Strains with a thyX deletion could not be obtained (PubMed:22034487). {ECO:0000269|PubMed:12657046, ECO:0000269|PubMed:22034487}. |
| Miscellaneous: | |
Was identified as a high-confidence drug target. {ECO:0000305|PubMed:19099550}. |
| Miscellaneous: | |
Crystallographic studies have shown that NADPH/NADP(+) binding expels both FAD and dUMP from the active site, by competing for the binding site (PubMed:16730023). However, the location of NADPH binding might not be biologically relevant (PubMed:18192395). {ECO:0000269|PubMed:16730023, ECO:0000305|PubMed:18192395}. |
| Similarity: | |
Belongs to the thymidylate synthase ThyX family. {ECO:0000255|HAMAP-Rule:MF_01408}. |
Annotations taken from UniProtKB at the EBI.