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Literature summary extracted from

  • Jourden, M.J.; Geiss, P.R.; Thomenius, M.J.; Horst, L.A.; Barty, M.M.; Brym, M.J.; Mulligan, G.B.; Almeida, R.M.; Kersteen, B.A.; Myers, N.R.; Snider, M.J.; Borders, C.L., Jr.; Edmiston, P.L.
    Transition state stabilization by six arginines clustered in the active site of creatine kinase (2005), Biochim. Biophys. Acta, 1751, 178-183.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.3.2 expression of soluble wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Oryctolagus cuniculus

Protein Variants

EC Number Protein Variants Comment Organism
2.7.3.2 R129A site-directed mutagenesis, inactive mutant Oryctolagus cuniculus
2.7.3.2 R129K site-directed mutagenesis, very highly reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
2.7.3.2 R131A site-directed mutagenesis, inactive mutant Oryctolagus cuniculus
2.7.3.2 R131K site-directed mutagenesis, reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
2.7.3.2 R235A site-directed mutagenesis, inactive mutant Oryctolagus cuniculus
2.7.3.2 R235K site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
2.7.3.2 R291K site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
2.7.3.2 R319K site-directed mutagenesis, reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
2.7.3.2 R340A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
2.7.3.2 R340K site-directed mutagenesis, reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
2.7.3.2 R340Q site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Oryctolagus cuniculus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.3.2 additional information
-
 additional information kinetics of wild-type and mutant enzymes Oryctolagus cuniculus
2.7.3.2 0.7
-
 ATP pH 9.0, 30°C, recombinant wild-type enzyme Oryctolagus cuniculus
2.7.3.2 0.7
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 1.1
-
 ATP pH 9.0, 30°C, recombinant mutant R340K Oryctolagus cuniculus
2.7.3.2 1.1
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 1.6
-
 ATP pH 9.0, 30°C, recombinant mutant R291K Oryctolagus cuniculus
2.7.3.2 1.6
-
 ATP pH 9.0, 30°C, recombinant mutant R340A Oryctolagus cuniculus
2.7.3.2 1.6
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 3.6
-
 ATP pH 9.0, 30°C, recombinant mutant R235K Oryctolagus cuniculus
2.7.3.2 3.6
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 7.3
-
 ATP pH 9.0, 30°C, recombinant mutant R340Q Oryctolagus cuniculus
2.7.3.2 7.3
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 8.2
-
 Creatine pH 9.0, 30°C, recombinant mutant R340A Oryctolagus cuniculus
2.7.3.2 9
-
 Creatine pH 9.0, 30°C, recombinant wild-type enzyme Oryctolagus cuniculus
2.7.3.2 10.3
-
 ATP pH 9.0, 30°C, recombinant mutant R129A Oryctolagus cuniculus
2.7.3.2 10.3
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 19
-
 Creatine pH 9.0, 30°C, recombinant mutant R340K Oryctolagus cuniculus
2.7.3.2 72
-
 Creatine pH 9.0, 30°C, recombinant mutant R235K Oryctolagus cuniculus
2.7.3.2 76
-
 Creatine pH 9.0, 30°C, recombinant mutant R291K Oryctolagus cuniculus
2.7.3.2 163
-
 Creatine pH 9.0, 30°C, recombinant mutant R340Q Oryctolagus cuniculus
2.7.3.2 167
-
 Creatine pH 9.0, 30°C, recombinant mutant R129A Oryctolagus cuniculus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.3.2 Mg2+ MgATP2-, at 1 mM, dissociation constants of wild-type and mutant enzymes Oryctolagus cuniculus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.3.2 ATP + creatine Oryctolagus cuniculus key enzyme in energy homeostasis ADP + phosphocreatine
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.3.2 Oryctolagus cuniculus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.3.2 recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) Oryctolagus cuniculus

Reaction

EC Number Reaction Comment Organism Reaction ID
2.7.3.2 ATP + creatine = ADP + phosphocreatine bireactant catalytic mechanism, transition state stabilization by six arginines clustered in the active site of the enzyme Oryctolagus cuniculus
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.3.2 skeletal muscle
-
 Oryctolagus cuniculus
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.3.2 ATP + creatine
-
 Oryctolagus cuniculus ADP + phosphocreatine
-
 r
2.7.3.2 ATP + creatine key enzyme in energy homeostasis Oryctolagus cuniculus ADP + phosphocreatine
-
 r

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.3.2 30
-
 assay at Oryctolagus cuniculus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.3.2 0.08
-
 ATP pH 9.0, 30°C, recombinant mutant R129A Oryctolagus cuniculus
2.7.3.2 0.08
-
 Creatine pH 9.0, 30°C, recombinant mutant R129A Oryctolagus cuniculus
2.7.3.2 0.08
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 0.54
-
 ATP pH 9.0, 30°C, recombinant mutant R235K Oryctolagus cuniculus
2.7.3.2 0.54
-
 Creatine pH 9.0, 30°C, recombinant mutant R235K Oryctolagus cuniculus
2.7.3.2 0.54
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 1.33
-
 ATP pH 9.0, 30°C, recombinant mutant R340Q Oryctolagus cuniculus
2.7.3.2 1.33
-
 Creatine pH 9.0, 30°C, recombinant mutant R340Q Oryctolagus cuniculus
2.7.3.2 1.33
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 2.14
-
 ATP pH 9.0, 30°C, recombinant mutant R340A Oryctolagus cuniculus
2.7.3.2 2.14
-
 Creatine pH 9.0, 30°C, recombinant mutant R340A Oryctolagus cuniculus
2.7.3.2 2.14
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 2.18
-
 ATP pH 9.0, 30°C, recombinant mutant R291K Oryctolagus cuniculus
2.7.3.2 2.18
-
 Creatine pH 9.0, 30°C, recombinant mutant R291K Oryctolagus cuniculus
2.7.3.2 2.18
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 50.1
-
 ATP pH 9.0, 30°C, recombinant mutant R340K Oryctolagus cuniculus
2.7.3.2 50.1
-
 Creatine pH 9.0, 30°C, recombinant mutant R340K Oryctolagus cuniculus
2.7.3.2 50.1
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus
2.7.3.2 148
-
 ATP pH 9.0, 30°C, recombinant wild-type enzyme Oryctolagus cuniculus
2.7.3.2 148
-
 Creatine pH 9.0, 30°C, recombinant wild-type enzyme Oryctolagus cuniculus
2.7.3.2 148
-
 ATP ATP in form of MgATP2- Oryctolagus cuniculus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.3.2 9
-
 assay at Oryctolagus cuniculus

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.3.2 ADP MgADP2- Oryctolagus cuniculus
2.7.3.2 ATP MgATP2- Oryctolagus cuniculus