Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Graziani, S.; Bernauer, J.; Skouloubris, S.; Graille, M.; Zhou, C.Z.; Marchand, C.; Decottignies, P.; van Tilbeurgh, H.; Myllykallio, H.; Liebl, U.
    Catalytic mechanism and structure of viral flavin-dependent thymidylate synthase ThyX (2006), J. Biol. Chem., 281, 24048-24057.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.148 wild-type and mutant ThyX proteins expressed in either Escherichia coli DH5alpha (deltathyA) or BL21 Paramecium bursaria Chlorella virus-1

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.148 crystal structure of the ThyX protein complexed to its FAD cofactor and solved by molecular replacement, to 2.3 A resolution, crystals belong to the P21212 space group with predicted two molecules per asymmetric unit and a solvent content of 51% Paramecium bursaria Chlorella virus-1

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.148 E190G does not confer thymidine-independent growth to an Escherichia coli strain lacking thymidylate synthase ThyA, is capable of binding FAD at a wild-type level, but lacks detectable oxidation and deprotonation Paramecium bursaria Chlorella virus-1
2.1.1.148 H177K does not confer thymidine-independent growth to an Escherichia coli strain lacking thymidylate synthase ThyA, 9.5% oxidation activity Paramecium bursaria Chlorella virus-1
2.1.1.148 H177Q confers thymidine-independent growth to an Escherichia coli strain lacking thymidylate synthase ThyA, 31% oxidation activity Paramecium bursaria Chlorella virus-1
2.1.1.148 H53K does not confer thymidine-independent growth to an Escherichia coli strain lacking thymidylate synthase ThyA, produces insoluble protein Paramecium bursaria Chlorella virus-1
2.1.1.148 H53Q does not confer thymidine-independent growth to an Escherichia coli strain lacking thymidylate synthase ThyA, produces insoluble protein Paramecium bursaria Chlorella virus-1
2.1.1.148 H79K does not confer thymidine-independent growth to an Escherichia coli strain lacking thymidylate synthase ThyA Paramecium bursaria Chlorella virus-1
2.1.1.148 H79Q confers thymidine-independent growth to an Escherichia coli strain lacking thymidylate synthase ThyA, 94% oxidation activity Paramecium bursaria Chlorella virus-1
2.1.1.148 R182A does not confer thymidine-independent growth to an Escherichia coli strain lacking thymidylate synthase ThyA, does not copurify with oxidized FAD, but is able to oxidize NADPH in the presence of 0.4 mM FAD Paramecium bursaria Chlorella virus-1
2.1.1.148 R90A does not confer thymidine-independent growth to an Escherichia coli strain lacking thymidylate synthase ThyA, looses 44% of its oxidation activity and shows no measurable deprotonation activity Paramecium bursaria Chlorella virus-1

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.148 5,10-methylenetetrahydrofolate
-
 Paramecium bursaria Chlorella virus-1
2.1.1.148 diethyldicarbonate decreases ThyX deprotonation activity at least 20fold, is partially reversible with hydroxylamine treatment Paramecium bursaria Chlorella virus-1

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.148 Paramecium bursaria Chlorella virus-1
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.148 by nickel chromatography and gel filtration, more than 95% pure Paramecium bursaria Chlorella virus-1

Storage Stability

EC Number Storage Stability Organism
2.1.1.148 -80°C, 50 mM HEPES buffer, pH 7.0, 10% glycerol Paramecium bursaria Chlorella virus-1

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.148 5,10-methylenetetrahydrofolate + dUMP + FADH2
-
 Paramecium bursaria Chlorella virus-1 dTMP + tetrahydrofolate + FAD
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.148 homotetramer crystallographic studies Paramecium bursaria Chlorella virus-1

Synonyms

EC Number Synonyms Comment Organism
2.1.1.148 flavin dependent thymidylate synthase
-
 Paramecium bursaria Chlorella virus-1
2.1.1.148 ThyX
-
 Paramecium bursaria Chlorella virus-1

Cofactor

EC Number Cofactor Comment Organism Structure
2.1.1.148 FAD
-
 Paramecium bursaria Chlorella virus-1
2.1.1.148 NADPH residue His53 is crucial for NADPH oxidation, is located in the vicinity of the redox active N-5 atom of the FAD ring system Paramecium bursaria Chlorella virus-1

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.1.1.148 0.016
-
 wild-type Paramecium bursaria Chlorella virus-1 5,10-methylenetetrahydrofolate
2.1.1.148 0.048
-
 R182A mutant Paramecium bursaria Chlorella virus-1 5,10-methylenetetrahydrofolate