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Literature summary extracted from

  • Feng, S.; Gong, Y.; Adilijiang, G.; Deng, H.
    Effects of the Fc-III tag on activity and stability of green fluorescent protein and human muscle creatine kinase (2013), Protein Sci., 22, 1008-1015.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.3.2 recombinant expression of Fc-III-tagged enzyme and of His-tagged enzyme in Escherichia coli strain BL21 (DE3)-plysS, respectively, the Fc-III-tagged GFP-fusion muscle creatine kinase is used as a model system to investigate effects of the Fc-III tag on activities and stabilities of recombinantly expressed multicysteine-containing proteins Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.7.3.2 additional information construction of a Fc-III-tagged GFP fusion muscle creatine kinase (CK) as a model system to investigate effects of the Fc-III tag on activities and stabilities of recombinantly expressed multicysteine-containing proteins. The Fc-III tag has no adverse effects on the fluorescence of GFP and reduces the occurrence of GFP misfolding due to incorrect Cys oxidation compared with the His-tagged protein. Activity and stability of the Fc-III-tagged creatine kinase is slightly lower than that of the tag-free creatine kinase, but is higher than that of the His-tagged creatine kinase as determined by the ratio of the oxidized versus reduced CK. A major portion of His-tagged CK is in its oxidized form, while that of the Fc-III-tagged CK is in its reduced form. A folding model of CK with different tags is proposed, overview. The decrease of the activity of CK may attribute to the tag-induced changes of the tertiary structures of the targeted proteins Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.3.2 guanidinium hydrochloride 0.1-3.0 M, under both conditions, the tag-free enzyme shows the lowest degree of aggregation, followed by His-tagged CK, and Fc-III-tagged CK has the highest degree of aggregation Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.3.2 Mg2+ required Homo sapiens

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 Homo sapiens
-
 ADP + phosphocreatine
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.3.2 Homo sapiens P06732
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.3.2 recombinant Fc-III-tagged enzyme from Escherichia coli strain BL21 (DE3)-plysS by IgG-Fc immunoaffinity chromatography, recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3)-plysS by nickel affinity chromatography Homo sapiens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.3.2 skeletal muscle
-
 Homo sapiens
-

Substrates and Products (Substrate)

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

Subunits

EC Number Subunits Comment Organism
2.7.3.2 More formation of the disulfide bond between the C74 and the C146 residues in the oxidized form Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.3.2 25
-
 assay at Homo sapiens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.3.2 52
-
 the recombinant enzymes are stable upto 50°C, the recombinant His-tagged and Fc-III-tagged enzymes show similar heat-resistance and are less stable than that of the tag-free enzyme. They start to lose their activities at 52°C and are relatively less active at the same denaturing temperature than tag-free enzyme. The DELTAT0.5 value of the tag-free enzyme is about 58°C and that of the tagged enzyme is around 57°C Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.3.2 9
-
 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.3.2 ATP
-
 Homo sapiens

General Information

EC Number General Information Comment Organism
2.7.3.2 additional information formation of the disulfide bond between the C74 and the C146 residues in the oxidized form Homo sapiens