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Literature summary for 1.1.1.267 extracted from

  • Henriksson, L.M.; Bjoerkelid, C.; Mowbray, S.L.; Unge, T.
    The 1.9 A resolution structure of Mycobacterium tuberculosis 1-deoxy-D-xylulose 5-phosphate reductoisomerase, a potential drug target (2006), Acta Crystallogr. Sect. D, D62, 807-813.
    View publication on PubMed

Application

Application Comment Organism
drug development the enzyme is an excellent drug target in a number of pathogenic organisms Mycobacterium tuberculosis

Cloned(Commentary)

Cloned (Comment) Organism
gene Rv2870c, expression of His-tagged enzyme in Escherichia coli strain BL21 Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged enzyme with inhibitor fosmidomycin, microbatch method, sitting drops mixed from 0.001 ml of protein solution, containing 5 mg/ml protein in 150 mM NaCl and 20 mM Tris–HCl pH 7.5, and 0.001 ml of crystallization buffer, containing 40% ethylene glycol, 2 mM MgSO4, 20 mM DTT, 0.2 mM EDTA and 100 mM acetate pH 5.0, streak-seeded immediately with a horse hair, 4 weeks, stabilization prior to flash-cooling in reservoir solution complemented with additional 10% ethylene glycol and 1 mM fosmidomycin, X-ray diffraction structure determination and analysis at 1.9 A resolution, molecular replacement Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
fosmidomycin
-
Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0072
-
NADPH
-
Mycobacterium tuberculosis
0.34
-
1-deoxy-D-xylulose 5-phosphate
-
Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ required Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1-deoxy-D-xylulose 5-phosphate + NADPH + H+ Mycobacterium tuberculosis second step of the deoxyxylulose 5-phosphate/methylerythritol 4-phosphate pathway with end product isopentenylphosphate 2-C-methyl-D-erythritol 4-phosphate + NADP+
-
?
1-deoxy-D-xylulose 5-phosphate + NADPH + H+ Mycobacterium tuberculosis H37Rv second step of the deoxyxylulose 5-phosphate/methylerythritol 4-phosphate pathway with end product isopentenylphosphate 2-C-methyl-D-erythritol 4-phosphate + NADP+
-
?

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
gene Rv2870c
-
Mycobacterium tuberculosis H37Rv
-
gene Rv2870c
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21 by metal ion affinity chromatography, and gel filtration, to 98% purity Mycobacterium tuberculosis

Reaction

Reaction Comment Organism Reaction ID
2-C-methyl-D-erythritol 4-phosphate + NADP+ = 1-deoxy-D-xylulose 5-phosphate + NADPH + H+ active site structure Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-deoxy-D-xylulose 5-phosphate + NADPH + H+
-
Mycobacterium tuberculosis 2-C-methyl-D-erythritol 4-phosphate + NADP+
-
?
1-deoxy-D-xylulose 5-phosphate + NADPH + H+ second step of the deoxyxylulose 5-phosphate/methylerythritol 4-phosphate pathway with end product isopentenylphosphate Mycobacterium tuberculosis 2-C-methyl-D-erythritol 4-phosphate + NADP+
-
?
1-deoxy-D-xylulose 5-phosphate + NADPH + H+
-
Mycobacterium tuberculosis H37Rv 2-C-methyl-D-erythritol 4-phosphate + NADP+
-
?
1-deoxy-D-xylulose 5-phosphate + NADPH + H+ second step of the deoxyxylulose 5-phosphate/methylerythritol 4-phosphate pathway with end product isopentenylphosphate Mycobacterium tuberculosis H37Rv 2-C-methyl-D-erythritol 4-phosphate + NADP+
-
?

Subunits

Subunits Comment Organism
dimer enzyme crystal structure, the dimer interface is created by interactions between the catalytic and connecting regions of each subunit, a sulfate is bound at the expected site of the phosphate moiety of the sugar substrate, molecular replacement structure analysis and comparison, overview Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
DXR
-
Mycobacterium tuberculosis
Rv2870c
-
Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
assay at Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.3
-
NADPH
-
Mycobacterium tuberculosis
5.3
-
1-deoxy-D-xylulose 5-phosphate
-
Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Mycobacterium tuberculosis