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Literature summary for 3.4.22.70 extracted from

  • Bentley, M.L.; Lamb, E.C.; McCafferty, D.G.
    Mutagenesis studies of substrate recognition and catalysis in the sortase A transpeptidase from Staphylococcus aureus (2008), J. Biol. Chem., 283, 14762-14771.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.22.70

Cloned(Commentary)

Cloned (Comment) Organism
mutant enzymes expressed in Escherichia coli Staphylococcus aureus

Protein Variants

Protein Variants Comment Organism
D170A Tm is 1.7°C higher than the Tm-value of wild-type enzyme. No change in kcat/Km compared to wild-type value Staphylococcus aureus
E171A Tm is 1.8°C lower than the Tm-value of wild-type enzyme. kcat/Km is 5.4fold lower than wild-type value Staphylococcus aureus
G167A Tm is 0.5°C lower than the Tm-value of wild-type enzyme. No change in kcat/KM compared to wild-type enzyme Staphylococcus aureus
L169A Tm is 1.8°C lower than the Tm-value of wild-type enzyme. kcat/Km is 93fold lower than wild-type value Staphylococcus aureus
Q172A Tm is 1.2°C lower than the Tm-value of wild-type enzyme. kcat/Km is 1.4fold lower than wild-type value Staphylococcus aureus
R197A Tm is 2.9°C lower than the Tm-value of wild-type enzyme. kcat/Km is 960fold lower than wild-type value Staphylococcus aureus
R197Cit generation of a semi-synthetic SrtA in which Arg197 is replaced with citrulline, a nonionizable analog. This change results in less than a 3fold decrease in kcat/KM, indicating that Arg197 utilizes a hydrogen bond, rather than an electrostatic interaction. Tm is 0.3°C higher than the Tm-value of wild-type enzyme Staphylococcus aureus
R197Cit kcat/Km is 5.9fold lower than wild-type value Staphylococcus aureus
R197K Tm is 2°C lower than the Tm-value of wild-type enzyme. kcat/Km is 690fold lower than wild-type value Staphylococcus aureus
V168A Tm is 1.7°C lower than the Tm-value of wild-type enzyme. kcat/Km is 5.5fold lower than wild-type value Staphylococcus aureus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.69
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme R197A Staphylococcus aureus
6.56
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme V168A Staphylococcus aureus
6.74
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme E171A Staphylococcus aureus
8.13
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme D170A Staphylococcus aureus
8.7
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme G167A Staphylococcus aureus
8.76
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, wild-type enzyme Staphylococcus aureus
9.14
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme L169A Staphylococcus aureus
10.4
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme R197K Staphylococcus aureus
12.7
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme Q172A Staphylococcus aureus
13.7
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme R197Cit Staphylococcus aureus

Localization

Localization Comment Organism GeneOntology No. Textmining

Organism

Organism UniProt Comment Textmining
Staphylococcus aureus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Staphylococcus aureus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Abz-LPETGG-Dap(Dnp)-NH2 + GGGGG
-
 Staphylococcus aureus Abz-LPETGGGGGG + Gly-Dap(Dnp)-NH2
-
 ?

Synonyms

Synonyms Comment Organism
SrtA
-
 Staphylococcus aureus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.000628
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme R197A Staphylococcus aureus
0.0019
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme R197K Staphylococcus aureus
0.0123
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme L169A Staphylococcus aureus
0.15
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme V168A Staphylococcus aureus
0.16
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme E171A Staphylococcus aureus
0.29
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme R197Cit Staphylococcus aureus
1.09
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme D170A Staphylococcus aureus
1.1
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, wild-type enzyme Staphylococcus aureus
1.13
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme G167A Staphylococcus aureus
1.13
-
 aminobenzoyl-LPETGG-diaminopropionic acid(dinitrophenol)-NH2 37°C, pH 7.5, mutant enzyme Q172A Staphylococcus aureus