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Literature summary for 2.7.6.1 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
Search for more literature for 2.7.6.1

Activating Compound

Activating Compound Comment Organism Structure
phosphate the enzyme is dependent on phosphate for its activity. The optimal concentration ranges from 10 mM to 40 mM Mycobacterium tuberculosis
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)

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

General Stability

General Stability Organism
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

Inhibitors Comment Organism Structure
ADP strong inhibition Mycobacterium tuberculosis
Ca2+
-
 Mycobacterium tuberculosis
Cu2+
-
 Mycobacterium tuberculosis
Fe2+
-
 Mycobacterium tuberculosis
GDP the enzyme is weakly sensitive to GDP Mycobacterium tuberculosis
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
phosphate
-
 Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
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

Localization Comment Organism GeneOntology No. Textmining

Metals/Ions

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-
Mycobacterium tuberculosis Rv1017
-
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
phosphoribosylpyrophosphate synthetase
-
 Mycobacterium tuberculosis
PRPPase
-
 Mycobacterium tuberculosis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
34.6
-
 Mg-ATP2- in the presence of 5 mM Mg2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
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
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
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
44.3
-
 Mg-ATP2- in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
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
45.1
-
 Mn-ATP2- in the presence of 5 mM Mn2+, in 50 mM potassium phosphate pH 8.0, at 37°C Mycobacterium tuberculosis
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

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

pH Range

pH Minimum pH Maximum Comment Organism
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

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

General Information

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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
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
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
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