Literature summary for 2.1.1.148 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.

Search for more literature for 2.1.1.148

Application

Application	Comment	Organism
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)

Cloned (Comment)	Organism
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)

Crystallization (Comment)	Organism
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

Protein Variants	Comment	Organism
H69E	fails to complement the Escherichia coli chi2913 cells	Mycobacterium tuberculosis
I65M	no impaired enzyme activity, encodes proteins supporting the growth of Escherichia coli chi2913 strain	Mycobacterium tuberculosis
I65M/L175M	produces active ThyX enzyme	Mycobacterium tuberculosis
K165A	fails to complement the Escherichia coli chi2913 cells	Mycobacterium tuberculosis
L175M	no impaired enzyme activity, encodes proteins supporting the growth of Escherichia coli chi2913 strain	Mycobacterium tuberculosis
R168A	fails to complement the Escherichia coli chi2913 cells	Mycobacterium tuberculosis
R95A	fails to complement the Escherichia coli chi2913 cells	Mycobacterium tuberculosis
R95D	fails to complement the Escherichia coli chi2913 cells	Mycobacterium tuberculosis
R95K	supports growth of Escherichia coli chi2913 cells	Mycobacterium tuberculosis
S105E	fails to complement the Escherichia coli chi2913 cells	Mycobacterium tuberculosis
Y108F	complemens the growth of Escherichia coli chi2913 cells	Mycobacterium tuberculosis

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
on Ni-NTA column	Mycobacterium tuberculosis

Storage Stability

Storage Stability	Organism
-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)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5,10-methylenetetrahydrofolate + BrdUMP + FADH2	-	Mycobacterium tuberculosis	?	-	?

Subunits

Subunits	Comment	Organism
homotetramer	crystallographic studies	Mycobacterium tuberculosis

Synonyms

Synonyms	Comment	Organism
flavin dependent thymidylate synthase	-	Mycobacterium tuberculosis
ThyX	-	Mycobacterium tuberculosis

Cofactor

Cofactor	Comment	Organism	Structure
FAD	-	Mycobacterium tuberculosis

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Release 2023.1 (January 2023)