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Literature summary for 2.7.1.60 extracted from

  • Coombes, D.; Davies, J.S.; Newton-Vesty, M.C.; Horne, C.R.; Setty, T.G.; Subramanian, R.; Moir, J.W.B.; Friemann, R.; Panjikar, S.; Griffin, M.D.W.; North, R.A.; Dobson, R.C.J.
    The basis for non-canonical ROK family function in the N-acetylmannosamine kinase from the pathogen Staphylococcus aureus (2020), J. Biol. Chem., 295, 3301-3315 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Staphylococcus aureus

Crystallization (Commentary)

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

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 [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 [Da] Molecular Weight Maximum [Da] Comment Organism
62000
-
sedimentation velocity analysis Staphylococcus aureus

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Staphylococcus aureus Q2G159
-
-
Staphylococcus aureus PS 47 Q2G159
-
-

Purification (Commentary)

Purification (Comment) Organism
Q-Sepharose column chromatography, phenyl-Sepharose column chromatography and Superdex S200 gel filtration Staphylococcus aureus

Substrates and Products (Substrate)

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

Subunits Comment Organism
homodimer small-angle X-ray scattering Staphylococcus aureus

Synonyms

Synonyms Comment Organism
N-acetylmannosamine kinase
-
Staphylococcus aureus
N-acetylmannosamine-6-phosphate 2-epimerase
-
Staphylococcus aureus
nanK
-
Staphylococcus aureus

Turnover Number [1/s]

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 [mM/s]

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