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Literature summary extracted from

  • Fantz, C.; Shaw, D.; Jennings, W.; Forsthoefel, A.; Kitchens, M.; Phan, J.; Minor, W.; Lebioda, L.; Berger, F.G.; Spencer, H.T.
    Drug-resistant variants of Escherichia coli thymidylate synthase: effects of substitutions at Pro-254 (2000), Mol. Pharmacol., 57, 359-366.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.1.1.45 medicine
-
 Mus musculus
2.1.1.45 medicine human TS mutants have potential applications in gene therapy by protecting hematopoietic progenitors from TS inhibitor toxicity, drug-resistant variants can be used to decrease the myelosuppressive side effects of TS-directed anticancer agents or to select genetically modified cells in vivo, clincal trials introduce chemoprotecting genes into hematopoietic cells Homo sapiens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.45 mutant P254D cDNA cloned into SacII/XhoI sites of the HaMSV retroviral vector and transfected into the TS negative cell line RJK88.13 Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.45
-
 Escherichia coli
2.1.1.45 mutant P254S Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.45 0.001
-
 methylenetetrahydropteroylgluamate mutant P254S Escherichia coli
2.1.1.45 0.0011
-
 methylenetetrahydropteroylgluamate wild-type Escherichia coli
2.1.1.45 0.0012
-
 2'-deoxyuridylate wild-type Escherichia coli
2.1.1.45 0.0012
-
 methylenetetrahydropteroylgluamate mutant P254G Escherichia coli
2.1.1.45 0.0015
-
 2'-deoxyuridylate mutant P254G Escherichia coli
2.1.1.45 0.0018
-
 2'-deoxyuridylate mutant P254F Escherichia coli
2.1.1.45 0.0018
-
 methylenetetrahydropteroylgluamate mutant P254F Escherichia coli
2.1.1.45 0.0027
-
 2'-deoxyuridylate mutant P254S Escherichia coli
2.1.1.45 0.0034
-
 2'-deoxyuridylate mutant P254D Escherichia coli
2.1.1.45 0.0118
-
 methylenetetrahydropteroylgluamate mutant P254D Escherichia coli

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 Mus musculus
-
 dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP Homo sapiens
-
 dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP Escherichia coli
-
 dihydrofolate + dTMP
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.45 Escherichia coli P0A884
-
-
2.1.1.45 Homo sapiens
-
 human
-
2.1.1.45 Mus musculus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.45
-
 Escherichia coli
2.1.1.45 Pro-254 mutant enzymes Homo sapiens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.1.1.45 bone marrow
-
 Mus musculus
-
2.1.1.45 bone marrow
-
 Homo sapiens
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Mus musculus dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Mus musculus dihydrofolate + dTMP
-
 r
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Homo sapiens dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Homo sapiens dihydrofolate + dTMP
-
 r
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Escherichia coli dihydrofolate + dTMP
-
 ?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
 Escherichia coli dihydrofolate + dTMP
-
 r
2.1.1.45 methylenetetrahydropteroylglutamate + dUMP
-
 Escherichia coli 7,8-dihydropteroylglutamate + thymidine phosphate
-
 r

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.1.1.45 37
-
 half-life of wild-type enzyme is 90 min, half-life of mutants P254F, P254S and P254D are approximately 50 min Homo sapiens