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Literature summary for 2.7.1.1 extracted from

  • Sun, S.
    Biochemical characterization of MODY2 glucokinase variants V62M and G72R reveals reduced enzymatic activities relative to wild-type (2009), Biochemistry, 48, 2514-2521.
    View publication on PubMed
Search for more literature for 2.7.1.1

Cloned(Commentary)

Cloned (Comment) Organism
wild type, mutant V62M,and G72R recombinant full-length human beta-cell glucokinases are expressed in Escherichia coli BL21(DE3) cells Homo sapiens

Protein Variants

Protein Variants Comment Organism
G72R mutant enzymes shows reduced rates of turnover (20% relative activity at 30°C, 10% relative activity at 37°C) and reduced glucose affinity relative to wild type, the mutant loses the stabilizing protein interactions with glucokinase regulatory protein, which may contribute to lower activity in vivo, the mutation associates with hyperglycemia in humans Homo sapiens
V62M mutant enzymes shows reduced rates of turnover (70% relative activity at 30°C, 40% relative activity at 37°C) and reduced glucose affinity relative to wild type, the mutant loses the stabilizing protein interactions with glucokinase regulatory protein, which may contribute to lower activity in vivo, the mutation associates with hyperglycemia in humans Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.7
-
 ATP wild type enzyme, at 30°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
0.77
-
 ATP wild type enzyme, at 37°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
0.82
-
 ATP mutant enzyme V62M, at 30°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
0.85
-
 ATP mutant enzyme V62M, at 37°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
1.1
-
 ATP mutant enzyme G72R, at 30°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
1.1
-
 ATP mutant enzyme G72R, at 37°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
52000
-
 x * 52000, SDS-PAGE Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens P35557
-
-

Purification (Commentary)

Purification (Comment) Organism
phenyl-Sepharose column chromatography, ammonium sulfate precipitation, Q-Sepharose column chromatography, glutathione-Sepharose 4B column chromatography Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + D-glucose
-
 Homo sapiens ADP + D-glucose 6-phosphate
-
 ?

Subunits

Subunits Comment Organism
? x * 52000, SDS-PAGE Homo sapiens

Synonyms

Synonyms Comment Organism
MODY2 glucokinase
-
 Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
48 52 half-life of 5-7 min at 50°C, the transition temperatures for the wild type enzyme are 48.4°C and 51.6°C in the absence and presence of saturating glucose (100 mM), respectively Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
24
-
 D-glucose mutant enzyme G72R, at 30°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
34
-
 D-glucose mutant enzyme G72R, at 37°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
54
-
 D-glucose mutant enzyme V62M, at 30°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
61
-
 D-glucose mutant enzyme V62M, at 37°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
74
-
 D-glucose wild type enzyme, at 30°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens
101
-
 D-glucose wild type enzyme, at 37°C, in 100 mM Tris and 125 mM KCl, pH 7.4, in the presence of 14 mM beta-mercaptoethanol or 5 mM dithiothreitol Homo sapiens

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Homo sapiens