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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.
    View publication on PubMed

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