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Literature summary extracted from

  • Lucarelli, A.P.; Buroni, S.; Pasca, M.R.; Rizzi, M.; Cavagnino, A.; Valentini, G.; Riccardi, G.; Chiarelli, L.R.
    Mycobacterium tuberculosis phosphoribosylpyrophosphate synthetase: biochemical features of a crucial enzyme for mycobacterial cell wall biosynthesis (2010), PLoS ONE, 5, e15494.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.6.1 phosphate the enzyme is dependent on phosphate for its activity. The optimal concentration ranges from 10 mM to 40 mM Mycobacterium tuberculosis
2.7.6.1 SO42- sulfate ions are able to stimulate the enzyme activity, but with respect to phosphate, are less effective. The optimal concentration ranges from 40 mM to 60 mM Mycobacterium tuberculosis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.6.1 expressed in Escherichia coli BL21(DE3)pLysS cells Mycobacterium tuberculosis

General Stability

EC Number General Stability Organism
2.7.6.1 dialysis against buffers such as 50 mM Tris-HCl, pH 8.0 or 50 mM HEPES-NaOH, pH 8.0 without phosphate results in a protein precipitation and complete loss of activity. The addition of 50 mM ammonium sulfate or 5 mM Mg-ATP to Tris-HCl, pH 8.0 allows the enzyme to preserve 20% of initial activity after a period of 16 h, whereas full activity is maintained with the addition of 50 mM phosphate Mycobacterium tuberculosis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.6.1 ADP strong inhibition Mycobacterium tuberculosis
2.7.6.1 Ca2+
-
 Mycobacterium tuberculosis
2.7.6.1 Cu2+
-
 Mycobacterium tuberculosis
2.7.6.1 Fe2+
-
 Mycobacterium tuberculosis
2.7.6.1 GDP the enzyme is weakly sensitive to GDP Mycobacterium tuberculosis
2.7.6.1 additional information no inhibitory effects are shown by the presence of pyrimidine nucleoside mono- or diphosphates or of histidine, up to 2 mM Mycobacterium tuberculosis
2.7.6.1 phosphate
-
 Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.6.1 0.06
-
 D-ribose 5-phosphate in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 0.07
-
 D-ribose 5-phosphate in the presence of 5 mM Mg2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 0.071
-
 D-ribose 5-phosphate in the absence of Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.6.1 Mg2+ allosterically activated by Mg2+ Mycobacterium tuberculosis
2.7.6.1 Mn2+ allosterically activated by Mn2+ Mycobacterium tuberculosis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.6.1 35000
-
 6 * 35000, SDS-PAGE Mycobacterium tuberculosis
2.7.6.1 220000
-
 gel filtration Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
2.7.6.1 Mycobacterium tuberculosis
-
-
-
2.7.6.1 Mycobacterium tuberculosis Rv1017
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.6.1 His-Trap HP column chromatography and Superdex 200 gel filtration Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.6.1 Mg-ATP2- + D-ribose 5-phosphate
-
 Mycobacterium tuberculosis Mg-AMP2- + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?
2.7.6.1 Mg-ATP2- + D-ribose 5-phosphate
-
 Mycobacterium tuberculosis Rv1017 Mg-AMP2- + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?
2.7.6.1 Mn-ATP2- + D-ribose 5-phosphate
-
 Mycobacterium tuberculosis Mn-AMP + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?
2.7.6.1 Mn-ATP2- + D-ribose 5-phosphate
-
 Mycobacterium tuberculosis Rv1017 Mn-AMP + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.6.1 homohexamer 6 * 35000, SDS-PAGE Mycobacterium tuberculosis

Synonyms

EC Number Synonyms Comment Organism
2.7.6.1 phosphoribosylpyrophosphate synthetase
-
 Mycobacterium tuberculosis
2.7.6.1 PRPPase
-
 Mycobacterium tuberculosis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.6.1 62 74 a highly stable enzyme, losing 50% of its activity in 10 min of incubation at 62°C, and showing a Tm of 69.3°C (without ligands) to 74.5°C (in the presence of 5 mM Mg-ATP) Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.6.1 34.6
-
 Mg-ATP2- in the presence of 5 mM Mg2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 35.1
-
 D-ribose 5-phosphate in the presence of 5 mM Mg2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 35.5
-
 Mg-ATP2- in the absence of free Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 37
-
 D-ribose 5-phosphate in the absence of Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 44.3
-
 Mg-ATP2- in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 44.7
-
 D-ribose 5-phosphate in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 45.1
-
 Mn-ATP2- in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 46.3
-
 Mn-ATP2- in the absence of free Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.6.1 8
-
 the enzyme shows an optimum at a pH value close to 8.0 Mycobacterium tuberculosis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.6.1 7 9.5 the enzyme possesses nearly 70% of its maximal activity at pH 9.5. The activity at pH 7 is 57% of the maximal one at pH 8.0 Mycobacterium tuberculosis

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.7.6.1 0.4
-
 in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis GDP
2.7.6.1 5
-
 in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis GDP
2.7.6.1 100
-
 in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis phosphate

General Information

EC Number General Information Comment Organism
2.7.6.1 physiological function the enzyme is crucial for mycobacterial cell wall biosynthesis Mycobacterium tuberculosis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.6.1 501.4
-
 D-ribose 5-phosphate in the presence of 5 mM Mg2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 521
-
 D-ribose 5-phosphate in the absence of Mg2+ or Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
2.7.6.1 745
-
 D-ribose 5-phosphate in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis