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Literature summary for 2.1.1.45 extracted from

  • Gokhale, R.S.; Agarwalla, S.; Santi, D.V.; Balaram, P.
    Covalent reinforcement of a fragile region in the dimeric enzyme thymidylate synthase stabilizes the protein against chaotrope-induced unfolding (1996), Biochemistry, 35, 7150-7158.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
cloned and overexpressed in Escherichia coli Lacticaseibacillus casei
plasmid pSCTS9, synthetic TS gene in pUC18 Lacticaseibacillus casei

Crystallization (Commentary)

Crystallization (Comment) Organism
-
Lacticaseibacillus casei

Protein Variants

Protein Variants Comment Organism
T155C/E188C/C244T formation of 2 disulfide crosslinks across the subunit interface results in dramatic stabilization of the overall structure of the protein, retention of secondary structure at temperatures as high as 90°C, engineering of an intersubunit crosslink yields a fully active enzyme Lacticaseibacillus casei

Inhibitors

Inhibitors Comment Organism Structure
5-fluoro-2'-deoxyuridylic acid
-
Lacticaseibacillus casei

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
2 * 35000, SDS-PAGE Lacticaseibacillus casei
36620
-
calculated molecular mass based on the sequence Lacticaseibacillus casei
71000
-
analytical ultracentrifugation Lacticaseibacillus casei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5,10-methylenetetrahydrofolate + dUMP Lacticaseibacillus casei
-
dihydrofolate + dTMP
-
?
5,10-methylenetetrahydrofolate + dUMP Tequatrovirus T4
-
dihydrofolate + dTMP
-
?

Organism

Organism UniProt Comment Textmining
Lacticaseibacillus casei
-
-
-
Tequatrovirus T4
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,10-methylenetetrahydrofolate + dUMP
-
Lacticaseibacillus casei dihydrofolate + dTMP
-
?
5,10-methylenetetrahydrofolate + dUMP
-
Lacticaseibacillus casei dihydrofolate + dTMP
-
r
5,10-methylenetetrahydrofolate + dUMP
-
Tequatrovirus T4 dihydrofolate + dTMP
-
?
5,10-methylenetetrahydrofolate + dUMP
-
Tequatrovirus T4 dihydrofolate + dTMP
-
r

Subunits

Subunits Comment Organism
dimer
-
Tequatrovirus T4
dimer 2 * 35000, SDS-PAGE Lacticaseibacillus casei

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
mutant enzyme, 2 disulfide bridges across the dimer interface of the enzyme engineered Lacticaseibacillus casei

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 70 mutant enzyme, 2 disulfide bridges across the dimer interface of the enzyme engineered Lacticaseibacillus casei

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
52
-
wild-type enzyme, precipitates at 52°C Lacticaseibacillus casei
90
-
mutant enzyme remains soluble, enhancement of activity is observed up to 60°C, possesses appreciable enzymatic activity at 65°C and retains secondary structure even at 90°C Lacticaseibacillus casei

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000005
-
5-fluoro-2'-deoxyuridylic acid
-
Lacticaseibacillus casei