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Literature summary for 2.4.2.21 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
Search for more literature for 2.4.2.21

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Sporomusa ovata

Crystallization (Commentary)

Crystallization (Comment) Organism
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
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

Protein Variants Comment Organism
I321S specific activity with p-cresol and 5,6-dimethylbenzimidazole decreased Sporomusa ovata
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
M87Q/I321S specific activity with p-cresol and 5,6-dimethylbenzimidazole highly decreased Sporomusa ovata

KM Value [mM]

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

Organism

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,6-dimethylbenzimidazole + beta-nicotinate D-ribonucleotide
-
 Sporomusa ovata nicotinate + N1-(5-phospho-alpha-D-ribosyl)-5,6-dimethylbenzimidazole
-
 ?
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 ?
-
 ?
p-cresol + beta-nicotinate D-ribonucleotide
-
 Sporomusa ovata nicotinate + 4-hydroxyphenyl alpha-D-ribose 5-phosphate
-
 ?
phenol + beta-nicotinate D-ribonucleotide
-
 Sporomusa ovata nicotinate + phenyl alpha-D-ribose 5-phosphate
-
 ?

Subunits

Subunits Comment Organism
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

Synonyms Comment Organism
ArsAB
-
 Sporomusa ovata

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

kcat/KM [mM/s]

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