Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Staphylococcus aureus |
Crystallization (Comment) | Organism |
---|---|
ligand-free, N-acetylmannosamine-bound and substrate analog GlcNAc-bound enzyme, sitting drop vapor diffusion method, using 0.2 M sodium malonate dibasic monohydrate and 20% (w/v) PEG 3350 | Staphylococcus aureus |
Protein Variants | Comment | Organism |
---|---|---|
Y160A | the mutant has a higher enzyme specificity (kcat/Km) with GlcNAc than for N-acetylmannosamine | Staphylococcus aureus |
Y160F | the mutant has a higher enzyme specificity (kcat/Km) with GlcNAc than for N-acetylmannosamine | Staphylococcus aureus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.049 | - |
N-acetyl-D-glucosamine | mutant enzyme Y160F, at pH 8.5 and 25°C | Staphylococcus aureus | |
0.057 | - |
N-acetyl-D-glucosamine | mutant enzyme Y160A, at pH 8.5 and 25°C | Staphylococcus aureus | |
0.26 | - |
N-acetyl-D-glucosamine | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus | |
0.3 | - |
N-acetyl-D-mannosamine | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus | |
0.73 | - |
ATP | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus | |
1.27 | - |
N-acetyl-D-mannosamine | mutant enzyme Y160F, at pH 8.5 and 25°C | Staphylococcus aureus | |
1.34 | - |
N-acetyl-D-mannosamine | mutant enzyme Y160A, at pH 8.5 and 25°C | Staphylococcus aureus | |
15 | - |
D-glucose | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
62000 | - |
sedimentation velocity analysis | Staphylococcus aureus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + N-acetyl-D-mannosamine | Staphylococcus aureus | - |
ADP + N-acetyl-D-mannosamine 6-phosphate | - |
r | |
ATP + N-acetyl-D-mannosamine | Staphylococcus aureus PS 47 | - |
ADP + N-acetyl-D-mannosamine 6-phosphate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Staphylococcus aureus | Q2G159 | - |
- |
Staphylococcus aureus PS 47 | Q2G159 | - |
- |
Purification (Comment) | Organism |
---|---|
Q-Sepharose column chromatography, phenyl-Sepharose column chromatography and Superdex S200 gel filtration | Staphylococcus aureus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + D-glucose | - |
Staphylococcus aureus | ADP + D-glucose 6-phosphate | - |
r | |
ATP + D-glucose | - |
Staphylococcus aureus PS 47 | ADP + D-glucose 6-phosphate | - |
r | |
ATP + N-acetyl-D-glucosamine | - |
Staphylococcus aureus | ADP + N-acetyl-D-glucosamine 6-phosphate | - |
r | |
ATP + N-acetyl-D-glucosamine | - |
Staphylococcus aureus PS 47 | ADP + N-acetyl-D-glucosamine 6-phosphate | - |
r | |
ATP + N-acetyl-D-mannosamine | - |
Staphylococcus aureus | ADP + N-acetyl-D-mannosamine 6-phosphate | - |
r | |
ATP + N-acetyl-D-mannosamine | - |
Staphylococcus aureus PS 47 | ADP + N-acetyl-D-mannosamine 6-phosphate | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | small-angle X-ray scattering | Staphylococcus aureus |
Synonyms | Comment | Organism |
---|---|---|
N-acetylmannosamine kinase | - |
Staphylococcus aureus |
N-acetylmannosamine-6-phosphate 2-epimerase | - |
Staphylococcus aureus |
nanK | - |
Staphylococcus aureus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
17.8 | - |
N-acetyl-D-glucosamine | mutant enzyme Y160A, at pH 8.5 and 25°C | Staphylococcus aureus | |
19.1 | - |
N-acetyl-D-glucosamine | mutant enzyme Y160F, at pH 8.5 and 25°C | Staphylococcus aureus | |
23.4 | - |
N-acetyl-D-glucosamine | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus | |
60 | - |
D-glucose | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus | |
216 | - |
N-acetyl-D-mannosamine | mutant enzyme Y160F, at pH 8.5 and 25°C | Staphylococcus aureus | |
223 | - |
N-acetyl-D-mannosamine | mutant enzyme Y160A, at pH 8.5 and 25°C | Staphylococcus aureus | |
262.6 | - |
ATP | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus | |
262.6 | - |
N-acetyl-D-mannosamine | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4 | - |
D-glucose | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus | |
90 | - |
N-acetyl-D-glucosamine | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus | |
167 | - |
N-acetyl-D-mannosamine | mutant enzyme Y160A, at pH 8.5 and 25°C | Staphylococcus aureus | |
171 | - |
N-acetyl-D-mannosamine | mutant enzyme Y160F, at pH 8.5 and 25°C | Staphylococcus aureus | |
310 | - |
N-acetyl-D-glucosamine | mutant enzyme Y160A, at pH 8.5 and 25°C | Staphylococcus aureus | |
390 | - |
N-acetyl-D-glucosamine | mutant enzyme Y160F, at pH 8.5 and 25°C | Staphylococcus aureus | |
871 | - |
N-acetyl-D-mannosamine | wild type enzyme, at pH 8.5 and 25°C | Staphylococcus aureus |