Literature summary extracted from
Sampathkumar, P.; Turley, S.; Ulmer, J.E.; Rhie, H.G.; Sibley, C.H.; Hol, W.G.
Structure of the Mycobacterium tuberculosis flavin dependent thymidylate synthase (MtbThyX) at 2.0A resolution (2005), J. Mol. Biol., 352, 1091-1104.
Application
EC Number |
Application |
Comment |
Organism |
---|
2.1.1.148 |
additional information |
complements the Escherichia coli chi2913 strain that lacks its conventional TS activity, residues Lys165 and Arg168 play critical roles in ThyX activity, possibly by governing access to the carbon atom to be methylated of a totally buried substrate dUMP |
Mycobacterium tuberculosis |
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
2.1.1.148 |
into the pET24d vector, overexpressed in Escherichia coli chi2913 strain, plasmid containing the I65M/L175M double mutant expressed in Escherichia coli BL21 (DE3) pLysS strain |
Mycobacterium tuberculosis |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
2.1.1.148 |
by the sitting-drop vapor diffusion method, in the presence of the cofactor FAD and the substrate analog BrdUMP, at 2.0 A resolution |
Mycobacterium tuberculosis |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
2.1.1.148 |
H69E |
fails to complement the Escherichia coli chi2913 cells |
Mycobacterium tuberculosis |
2.1.1.148 |
I65M |
no impaired enzyme activity, encodes proteins supporting the growth of Escherichia coli chi2913 strain |
Mycobacterium tuberculosis |
2.1.1.148 |
I65M/L175M |
produces active ThyX enzyme |
Mycobacterium tuberculosis |
2.1.1.148 |
K165A |
fails to complement the Escherichia coli chi2913 cells |
Mycobacterium tuberculosis |
2.1.1.148 |
L175M |
no impaired enzyme activity, encodes proteins supporting the growth of Escherichia coli chi2913 strain |
Mycobacterium tuberculosis |
2.1.1.148 |
R168A |
fails to complement the Escherichia coli chi2913 cells |
Mycobacterium tuberculosis |
2.1.1.148 |
R95A |
fails to complement the Escherichia coli chi2913 cells |
Mycobacterium tuberculosis |
2.1.1.148 |
R95D |
fails to complement the Escherichia coli chi2913 cells |
Mycobacterium tuberculosis |
2.1.1.148 |
R95K |
supports growth of Escherichia coli chi2913 cells |
Mycobacterium tuberculosis |
2.1.1.148 |
S105E |
fails to complement the Escherichia coli chi2913 cells |
Mycobacterium tuberculosis |
2.1.1.148 |
Y108F |
complemens the growth of Escherichia coli chi2913 cells |
Mycobacterium tuberculosis |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
2.1.1.148 |
Mycobacterium tuberculosis |
- |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
2.1.1.148 |
on Ni-NTA column |
Mycobacterium tuberculosis |
Storage Stability
EC Number |
Storage Stability |
Organism |
---|
2.1.1.148 |
-80°C, 10 mM Tris-HCl buffer, pH 7.2, 50 mM NaCl, 2 mM DTT, 100 mM FAD |
Mycobacterium tuberculosis |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
2.1.1.148 |
5,10-methylenetetrahydrofolate + BrdUMP + FADH2 |
- |
Mycobacterium tuberculosis |
? |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
2.1.1.148 |
homotetramer |
crystallographic studies |
Mycobacterium tuberculosis |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
2.1.1.148 |
flavin dependent thymidylate synthase |
- |
Mycobacterium tuberculosis |
2.1.1.148 |
ThyX |
- |
Mycobacterium tuberculosis |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
2.1.1.148 |
FAD |
- |
Mycobacterium tuberculosis |
|