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Literature summary extracted from

  • Gibson, L.M.; Lovelace, L.L.; Lebioda, L.
    The R163K mutant of human thymidylate synthase is stabilized in an active conformation: structural asymmetry and reactivity of cysteine 195 (2008), Biochemistry, 47, 4636-4643.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.45 plasmid containing the mutation was transformed into the Escherichia coli TX61-(thyA-) bacterial strain that does not produce its own thymidylate synthase enzyme Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.45 grown in low-salt conditions, 100 mM Tris, pH 9.0, 20 mM beta-mercaptoethanol, 3 mM KH2PO4, and 10-20% PEG 4K, by hanging drop diffusion at 4 °C Homo sapiens
2.1.1.45 wild-type and mutant R163K. Data show that all subunits of the mutant are in the active conformation, while wild-type crystallizes as the inactive conformer. Structure reveals differences in the environment of catalytic residue Cys195 Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.45 R163K mutation results in destabilization of inactive conformation. Activity is about 33% higher than in wild-type. Crystallization data show that all subunits of the mutant are in the active conformation, while wild-type crystallizes as the inactive conformer. Structure reveals differences in the environment of catalytic residue Cys195 Homo sapiens
2.1.1.45 R163K the activity of this mutant is 33% higher than that of wt hTS Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP Homo sapiens
-
 7,8-dihydrofolate + dTMP
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.45 Homo sapiens P04818
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Homo sapiens 7,8-dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP mTHF, TS is the sole source of de novo thymidylate which is required for DNA synthesis Homo sapiens 7,8-dihydrofolate + dTMP
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.45 dimer asymmetric, X-ray diffraction Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
2.1.1.45 HTS
-
 Homo sapiens