Literature summary for 2.7.3.2 extracted from

Wu, Q.Y.; Wei, F.; Zhu, Y.Y.; Tong, Y.X.; Cao, J.; Zhou, P.; Li, Z.Y.; Zeng, L.Y.; Li, F.; Wang, X.Y.; Xu, K.L.
Roles of amino acid residues H66 and D326 in the creatine kinase activity and structural stability (2018), Int. J. Biol. Macromol., 107, 512-520 .

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21 cells	Homo sapiens
expressed in Escherichia coli BL21(DE3) cells	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D326A	the mutant displays obviously loss of activity, substrate synergism and stability as compared to the wild type enzyme	Homo sapiens
D326A	the mutant enzyme displays strong loss of activity, substrate synergism and stability with partially unfolded state and aggregation	Homo sapiens
D326E	the mutant displays obviously loss of activity, substrate synergism and stability as compared to the wild type enzyme	Homo sapiens
D326E	the mutant enzyme displays loss of activity	Homo sapiens
H66P	the mutant displays obviously loss of activity, substrate synergism and stability as compared to the wild type enzyme	Homo sapiens
H66P	the mutant enzyme displays strong loss of activity, substrate synergism and stability with partially unfolded state and aggregation	Homo sapiens
H66P/D326A	the mutant displays obviously loss of activity, substrate synergism and stability as compared to the wild type enzyme	Homo sapiens
H66P/D326A	the mutant enzyme displays severe loss of activity, substrate synergism and stability	Homo sapiens
H66R	the mutant displays obviously loss of activity, substrate synergism and stability as compared to the wild type enzyme	Homo sapiens
H66R	the mutant enzyme displays loss of activity	Homo sapiens

General Stability

General Stability	Organism
a His6-tag portion has no effect on the enzyme activity	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.56	-	ATP	wild type enzyme, at pH 8.0 and 25°C	Homo sapiens
0.56	-	ATP	recombinant wild type enzyme, at 25Â°C, pH not specified in the publication	Homo sapiens
1.14	-	ATP	recombinant mutant enzyme D326E, at 25Â°C, pH not specified in the publication	Homo sapiens
1.14	-	ATP	mutant enzyme D326E, at pH 8.0 and 25°C	Homo sapiens
1.4	-	ATP	recombinant mutant enzyme H66R, at 25Â°C, pH not specified in the publication	Homo sapiens
1.4	-	ATP	mutant enzyme H66R, at pH 8.0 and 25°C	Homo sapiens
3.98	-	ATP	recombinant mutant enzyme D326A, at 25Â°C, pH not specified in the publication	Homo sapiens
3.98	-	ATP	mutant enzyme D326A, at pH 8.0 and 25°C	Homo sapiens
4.98	-	ATP	recombinant mutant enzyme H66P, at 25Â°C, pH not specified in the publication	Homo sapiens
4.98	-	ATP	mutant enzyme H66P, at pH 8.0 and 25°C	Homo sapiens
8.96	-	ATP	recombinant mutant enzyme H66P/D326A, at 25Â°C, pH not specified in the publication	Homo sapiens
8.96	-	ATP	mutant enzyme H66P/D326A, at pH 8.0 and 25°C	Homo sapiens
9.15	-	Creatine	wild type enzyme, at pH 8.0 and 25°C	Homo sapiens
9.15	-	Creatine	recombinant wild type enzyme, at 25Â°C, pH not specified in the publication	Homo sapiens
19.28	-	Creatine	recombinant mutant enzyme D326E, at 25Â°C, pH not specified in the publication	Homo sapiens
19.28	-	Creatine	mutant enzyme D326E, at pH 8.0 and 25°C	Homo sapiens
24.11	-	Creatine	recombinant mutant enzyme H66R, at 25Â°C, pH not specified in the publication	Homo sapiens
24.11	-	Creatine	mutant enzyme H66R, at pH 8.0 and 25°C	Homo sapiens
79.88	-	Creatine	recombinant mutant enzyme D326A, at 25Â°C, pH not specified in the publication	Homo sapiens
79.88	-	Creatine	mutant enzyme D326A, at pH 8.0 and 25°C	Homo sapiens
87.12	-	Creatine	recombinant mutant enzyme H66P, at 25Â°C, pH not specified in the publication	Homo sapiens
87.12	-	Creatine	mutant enzyme H66P, at pH 8.0 and 25°C	Homo sapiens
156.44	-	Creatine	recombinant mutant enzyme H66P/D326A, at 25Â°C, pH not specified in the publication	Homo sapiens
156.44	-	Creatine	mutant enzyme H66P/D326A, at pH 8.0 and 25°C	Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
86000	-	gel filtration	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + creatine	Homo sapiens	-	ADP + phosphocreatine	-	?
ATP + creatine	Homo sapiens	-	ADP + phosphocreatine	-	r

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P06732	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Homo sapiens
affinity chromatography	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
muscle	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + creatine	-	Homo sapiens	ADP + phosphocreatine	-	?
ATP + creatine	-	Homo sapiens	ADP + phosphocreatine	-	r

Synonyms

Synonyms	Comment	Organism
M-CK	-	Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
25	60	the wild type enzyme remains stable after 10 min at 25-45Â°C. The activity drops to about 70% and after 10 min at 50Â°C. The enzyme is almost completely inactive after 10 min at 60Â°C	Homo sapiens
25	65	the activity of the wild type enzyme has few changes after heat treatment for 10 min at temperatures below 48°C. A steep decrease of activity is observed between 48°C and 60°C	Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
28.99	-	ATP	recombinant mutant enzyme H66P/D326A, at 25Â°C, pH not specified in the publication	Homo sapiens
28.99	-	Creatine	recombinant mutant enzyme H66P/D326A, at 25Â°C, pH not specified in the publication	Homo sapiens
28.99	-	Creatine	mutant enzyme H66P/D326A, at pH 8.0 and 25°C	Homo sapiens
62.15	-	ATP	recombinant mutant enzyme H66P, at 25Â°C, pH not specified in the publication	Homo sapiens
62.15	-	Creatine	recombinant mutant enzyme H66P, at 25Â°C, pH not specified in the publication	Homo sapiens
62.15	-	Creatine	mutant enzyme H66P, at pH 8.0 and 25°C	Homo sapiens
67.78	-	ATP	recombinant mutant enzyme D326A, at 25Â°C, pH not specified in the publication	Homo sapiens
67.78	-	Creatine	recombinant mutant enzyme D326A, at 25Â°C, pH not specified in the publication	Homo sapiens
67.78	-	Creatine	mutant enzyme D326A, at pH 8.0 and 25°C	Homo sapiens
136.21	-	ATP	recombinant mutant enzyme H66R, at 25Â°C, pH not specified in the publication	Homo sapiens
136.21	-	Creatine	recombinant mutant enzyme H66R, at 25Â°C, pH not specified in the publication	Homo sapiens
136.21	-	Creatine	mutant enzyme H66R, at pH 8.0 and 25°C	Homo sapiens
157.66	-	ATP	recombinant mutant enzyme D326E, at 25Â°C, pH not specified in the publication	Homo sapiens
157.66	-	Creatine	recombinant mutant enzyme D326E, at 25Â°C, pH not specified in the publication	Homo sapiens
157.66	-	Creatine	mutant enzyme D326E, at pH 8.0 and 25°C	Homo sapiens
177.66	-	Creatine	wild type enzyme, at pH 8.0 and 25°C	Homo sapiens
177.66	-	ATP	recombinant wild type enzyme, at 25Â°C, pH not specified in the publication	Homo sapiens
177.66	-	Creatine	recombinant wild type enzyme, at 25Â°C, pH not specified in the publication	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Homo sapiens