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Literature summary for 3.1.3.5 extracted from

  • Grobosky, C.L.; Lopez, J.B.; Rennie, S.; Skopelitis, D.J.; Wiest, A.T.; Bingman, C.A.; Bitto, E.
    Structural basis of substrate specificity and selectivity of murine cytosolic 5-nucleotidase III (2012), J. Mol. Biol., 423, 540-554.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
-
Mus musculus

Crystallization (Commentary)

Crystallization (Comment) Organism
X-ray structure of catalytically inactivate variant D51N of murine cN-III with substrate uridine 5'-monophosphate at 1.74 A resolution. The aromatic ring of a nitrogenous base of the substrate nucleotide is stabilized by parallel pi-stacking interactions with conserved aromatic rings of Trp113 and His68. The nitrogenous base is further stabilized by T-shaped stacking with the conserved aromatic ring of Tyr114, as well as by polar contacts with side chains of Thr66 and Ser117. Two water molecules help to stabilize the nucleotide binding by bridging it to protein residues Asp72 and His68 via hydrogen bonds. Glu96 is responsible for recognition of ribose ring via two hydrogen bonds Mus musculus

Protein Variants

Protein Variants Comment Organism
D51N complete loss of activity Mus musculus
E96S significant decrease in catalytic ability Mus musculus
H68A significant decrease in catalytic ability Mus musculus
N69G slightly higher Km compared to the wild?type Mus musculus
S117G no significant changes compared to wild-type Mus musculus
T66G no significant changes compared to wild-type Mus musculus
W113A significant decrease in catalytic ability Mus musculus
Y114A significant decrease in catalytic ability Mus musculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1
-
5'-UMP wild-type, pH 7.5, 37°C Mus musculus
1.1
-
5'-UMP mutant S117G, pH 7.5, 37°C Mus musculus
1.1
-
5'-UMP mutant T66G, pH 7.5, 37°C Mus musculus
2.1
-
5'-UMP mutant N69G, pH 7.5, 37°C Mus musculus
2.3
-
5'-UMP mutant Y114A, pH 7.5, 37°C Mus musculus
3.2
-
5'-UMP mutant H68A, pH 7.5, 37°C Mus musculus
4.8
-
5'-UMP mutant E96S, pH 7.5, 37°C Mus musculus
29
-
5'-UMP mutant W11A, pH 7.5, 37°C Mus musculus

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Mus musculus 5829
-

Organism

Organism UniProt Comment Textmining
Mus musculus Q9D020
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5'-UMP + H2O
-
Mus musculus uridine + phosphate
-
?

Synonyms

Synonyms Comment Organism
cN-III
-
Mus musculus
NT5C3A
-
Mus musculus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.12
-
5'-UMP mutant H68A, pH 7.5, 37°C Mus musculus
0.14
-
5'-UMP mutant Y114A, pH 7.5, 37°C Mus musculus
0.29
-
5'-UMP mutant W11A, pH 7.5, 37°C Mus musculus
0.38
-
5'-UMP mutant E96S, pH 7.5, 37°C Mus musculus
1.1
-
5'-UMP mutant N69G, pH 7.5, 37°C Mus musculus
1.2
-
5'-UMP wild-type, pH 7.5, 37°C Mus musculus
1.7
-
5'-UMP mutant S117G, pH 7.5, 37°C Mus musculus
1.9
-
5'-UMP mutant T66G, pH 7.5, 37°C Mus musculus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.01
-
5'-UMP mutant W11A, pH 7.5, 37°C Mus musculus
0.037
-
5'-UMP mutant H68A, pH 7.5, 37°C Mus musculus
0.06
-
5'-UMP mutant Y114A, pH 7.5, 37°C Mus musculus
0.08
-
5'-UMP mutant E96S, pH 7.5, 37°C Mus musculus
0.55
-
5'-UMP mutant N69G, pH 7.5, 37°C Mus musculus
1.1
-
5'-UMP wild-type, pH 7.5, 37°C Mus musculus
1.5
-
5'-UMP mutant S117G, pH 7.5, 37°C Mus musculus
1.7
-
5'-UMP mutant T66G, pH 7.5, 37°C Mus musculus