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Literature summary extracted from

  • Muenchhoff, J.; Siddiqui, K.S.; Neilan, B.A.
    Identification of two residues essential for the stringent substrate specificity and active site stability of the prokaryotic L-arginine:glycine amidinotransferase CyrA (2012), FEBS J., 279, 805-815.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.4.1 gene cyrA, expression of N-terminal His6-tagged CyrA in Escherichia coli strain BL21(DE3) Cylindrospermopsis raciborskii

Protein Variants

EC Number Protein Variants Comment Organism
2.1.4.1 F245N site-directed mutagenesis, exchange analogously to the human L-arginine:glycine amidinotransferase, h-AGAT. F245N variant seems to lack the potential ligand-induced conformational changes. Higher activity of the F245N variant is enthalpy-driven Cylindrospermopsis raciborskii
2.1.4.1 F245N/S247M site-directed mutagenesis, exchange analogously to the human L-arginine:glycine amidinotransferase, h-AGAT Cylindrospermopsis raciborskii
2.1.4.1 additional information the residue replacements do not change the kinetic mechanism of the three variant enzymes. The mutant protein variants have broad substrate specificity, e.g. the F245N variant additionally accepts creatine, with 7% relative activity Cylindrospermopsis raciborskii
2.1.4.1 S247M site-directed mutagenesis, exchange analogously to the human L-arginine:glycine amidinotransferase, h-AGAT Cylindrospermopsis raciborskii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.4.1 50000
-
 x * 50000, about, SDS-PAGE Cylindrospermopsis raciborskii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.4.1 L-arginine + glycine Cylindrospermopsis raciborskii
-
 L-ornithine + guanidinoacetate
-
 ?
2.1.4.1 L-arginine + glycine Cylindrospermopsis raciborskii AWT205
-
 L-ornithine + guanidinoacetate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.4.1 Cylindrospermopsis raciborskii B0LI36 gene cyrA
-
2.1.4.1 Cylindrospermopsis raciborskii AWT205 B0LI36 gene cyrA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.4.1 recombinant wild-type and mutant N-terminal His6-tagged CyrAs from Escherichia coli strain BL21(DE3) by metal affinity chromatography to over 95% purity Cylindrospermopsis raciborskii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.4.1 guanidinoacetate + glycine
-
 Cylindrospermopsis raciborskii ?
-
 ?
2.1.4.1 guanidinoacetate + glycine
-
 Cylindrospermopsis raciborskii AWT205 ?
-
 ?
2.1.4.1 L-arginine + glycine
-
 Cylindrospermopsis raciborskii L-ornithine + guanidinoacetate
-
 ?
2.1.4.1 L-arginine + glycine
-
 Cylindrospermopsis raciborskii AWT205 L-ornithine + guanidinoacetate
-
 ?
2.1.4.1 additional information no activity with creatine, L-homoarginine, and L-canavanine by the wild-type enzyme, while mutants F245N, S247M, and F245N/S247M are also active L-homoarginine and L-canavanine, mutant F254N also slightly with creatine, overview Cylindrospermopsis raciborskii ?
-
 ?
2.1.4.1 additional information no activity with creatine, L-homoarginine, and L-canavanine by the wild-type enzyme, while mutants F245N, S247M, and F245N/S247M are also active L-homoarginine and L-canavanine, mutant F254N also slightly with creatine, overview Cylindrospermopsis raciborskii AWT205 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.4.1 ? x * 50000, about, SDS-PAGE Cylindrospermopsis raciborskii
2.1.4.1 More structure-function-stability relationship, overview Cylindrospermopsis raciborskii

Synonyms

EC Number Synonyms Comment Organism
2.1.4.1 CyrA
-
 Cylindrospermopsis raciborskii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.4.1 30
-
 recombinant mutant F245N Cylindrospermopsis raciborskii
2.1.4.1 35
-
 recombinant wild-type CyrA and mutant F245N/S247M Cylindrospermopsis raciborskii
2.1.4.1 40
-
 recombinant mutant S247M Cylindrospermopsis raciborskii

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.1.4.1 30 45 different optima for wild-type and mutant CyrAs, above 40°C activity decreases rapidly for all enzymes, including the S247M variant, which shows 18% of maximal activity at 45°C Cylindrospermopsis raciborskii

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.1.4.1 additional information
-
 the three mutant variants have lower Tmax values than the wild-type with 54°C compared to 48°C, 51.5°C and 49.5°C for F245N, S247M, and F245N/S247M, respectively Cylindrospermopsis raciborskii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.4.1 7
-
 recombinant mutant F245N/S247M Cylindrospermopsis raciborskii
2.1.4.1 8
-
 recombinant mutants F245N and S247M Cylindrospermopsis raciborskii
2.1.4.1 8.5
-
 recombinant wild-type CyrA Cylindrospermopsis raciborskii

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.1.4.1 7 9 above pH 9, the activity dropped rapidly for all enzymes Cylindrospermopsis raciborskii

General Information

EC Number General Information Comment Organism
2.1.4.1 additional information ligand-induced conformational changes in wild-type CyrA, structure-function-stability relationship, overview Cylindrospermopsis raciborskii