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Literature summary extracted from

  • Newmister, S.A.; Chan, C.H.; Escalante-Semerena, J.C.; Rayment, I.
    Structural Insights into the Function of the Nicotinate Mononucleotide:phenol/p-cresol Phosphoribosyltransferase (ArsAB) Enzyme from Sporomusa ovata (2012), Biochemistry, 51, 8571-8582.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.21 expressed in Escherichia coli Sporomusa ovata

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.2.21 phloroglucinol is identified from an additive screen as a compound that dramatically improves the size, stability, and diffraction properties of the crystals. Subsequent structural studies show that phloroglucinol binds in the active site of ArsA Sporomusa ovata
2.4.2.21 the three-dimensional crystal structure of Sporomusa ovata ArsAB in its substrate-free form (2.1 A), and in complex with DMB (1.5 A), p-cresol (2.1 A), phloroglucinol (1.6 A), and phenol (2.4 A) is reported. Only the active site in ArsA is occupied by substrate. Analysis of the ArsB active site identified an arginyl side chain that may block access to the site Sporomusa ovata

Protein Variants

EC Number Protein Variants Comment Organism
2.4.2.21 I321S specific activity with p-cresol and 5,6-dimethylbenzimidazole decreased Sporomusa ovata
2.4.2.21 M87Q effect of the M87Q mutation is most pronounced with respect to phenolic specificity, as the specific activity with p-cresol drops by 2.6fold while a 1.2fold increase in activity with 5,6-dimethylbenzimidazole is observed Sporomusa ovata
2.4.2.21 M87Q/I321S specific activity with p-cresol and 5,6-dimethylbenzimidazole highly decreased Sporomusa ovata

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.2.21 0.05
-
 phenol pH 9.0, 37°C, wild-type Sporomusa ovata
2.4.2.21 0.415
-
 5,6-Dimethylbenzimidazole pH 9.0, 37°C, wild-type Sporomusa ovata

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.21 Sporomusa ovata F6MZ55 and F6MZ56 subunit ArsA: F6MZ55, subunit ArsB: F6MZ56
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.2.21 0.0000007
-
 substrate p-cresol, pH 9.0, 37°C, mutant M87Q/I321S Sporomusa ovata
2.4.2.21 0.0000024
-
 substrate 5,6-dimethylbenzimidazole, pH 9.0, 37°C, mutant M87Q/I321S Sporomusa ovata
2.4.2.21 0.0000083
-
 substrate p-cresol, pH 9.0, 37°C, mutant I321S Sporomusa ovata
2.4.2.21 0.0000142
-
 substrate p-cresol, pH 9.0, 37°C, mutant M87Q Sporomusa ovata
2.4.2.21 0.0000167
-
 substrate 5,6-dimethylbenzimidazole, pH 9.0, 37°C, mutant I321S Sporomusa ovata
2.4.2.21 0.0000376
-
 substrate p-cresol, pH 9.0, 37°C, wild-type Sporomusa ovata
2.4.2.21 0.00016
-
 substrate 5,6-dimethylbenzimidazole, pH 9.0, 37°C, wild-type Sporomusa ovata
2.4.2.21 0.000192
-
 substrate 5,6-dimethylbenzimidazole, pH 9.0, 37°C, mutant M87Q Sporomusa ovata

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.21 5,6-dimethylbenzimidazole + beta-nicotinate D-ribonucleotide
-
 Sporomusa ovata nicotinate + N1-(5-phospho-alpha-D-ribosyl)-5,6-dimethylbenzimidazole
-
 ?
2.4.2.21 additional information phloroglucinol is identified from an additive screen as a compound that dramatically improves the size, stability, and diffraction properties of the crystals. Subsequent structural studies show that phloroglucinol binds in the active site of ArsA Sporomusa ovata ?
-
 ?
2.4.2.21 p-cresol + beta-nicotinate D-ribonucleotide
-
 Sporomusa ovata nicotinate + 4-hydroxyphenyl alpha-D-ribose 5-phosphate
-
 ?
2.4.2.21 phenol + beta-nicotinate D-ribonucleotide
-
 Sporomusa ovata nicotinate + phenyl alpha-D-ribose 5-phosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.2.21 heterodimer ArsA and ArsB. Only the active site in ArsA is occupied by substrate. Analysis of the ArsB active site identified an arginyl side chain that may block access to the site Sporomusa ovata

Synonyms

EC Number Synonyms Comment Organism
2.4.2.21 ArsAB
-
 Sporomusa ovata

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.21 37
-
 assay at Sporomusa ovata

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.2.21 0.4
-
 phenol pH 9.0, 37°C, wild-type Sporomusa ovata
2.4.2.21 1.6
-
 5,6-Dimethylbenzimidazole pH 9.0, 37°C, wild-type Sporomusa ovata

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.2.21 9
-
 assay at Sporomusa ovata

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.2.21 3.9
-
 5,6-Dimethylbenzimidazole pH 9.0, 37°C, wild-type Sporomusa ovata
2.4.2.21 8
-
 phenol pH 9.0, 37°C, wild-type Sporomusa ovata