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Literature summary for 2.1.4.1 extracted from
- 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.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene cyrA, expression of N-terminal His6-tagged CyrA in Escherichia coli strain BL21(DE3) | Cylindrospermopsis raciborskii |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| 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 |
| F245N/S247M | site-directed mutagenesis, exchange analogously to the human L-arginine:glycine amidinotransferase, h-AGAT | Cylindrospermopsis raciborskii |
| 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 |
| S247M | site-directed mutagenesis, exchange analogously to the human L-arginine:glycine amidinotransferase, h-AGAT | Cylindrospermopsis raciborskii |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 50000 | - |
x * 50000, about, SDS-PAGE | Cylindrospermopsis raciborskii |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-arginine + glycine | Cylindrospermopsis raciborskii | - |
L-ornithine + guanidinoacetate | - |
? |  |
| L-arginine + glycine | Cylindrospermopsis raciborskii AWT205 | - |
L-ornithine + guanidinoacetate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Cylindrospermopsis raciborskii | B0LI36 | gene cyrA | - |
| Cylindrospermopsis raciborskii AWT205 | B0LI36 | gene cyrA | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| 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)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| guanidinoacetate + glycine | - |
Cylindrospermopsis raciborskii | ? | - |
? | |
| guanidinoacetate + glycine | - |
Cylindrospermopsis raciborskii AWT205 | ? | - |
? | |
| L-arginine + glycine | - |
Cylindrospermopsis raciborskii | L-ornithine + guanidinoacetate | - |
? | |
| L-arginine + glycine | - |
Cylindrospermopsis raciborskii AWT205 | L-ornithine + guanidinoacetate | - |
? | |
| 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 | ? | - |
? | |
| 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
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 50000, about, SDS-PAGE | Cylindrospermopsis raciborskii |
| More | structure-function-stability relationship, overview | Cylindrospermopsis raciborskii |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CyrA | - |
Cylindrospermopsis raciborskii |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
recombinant mutant F245N | Cylindrospermopsis raciborskii |
| 35 | - |
recombinant wild-type CyrA and mutant F245N/S247M | Cylindrospermopsis raciborskii |
| 40 | - |
recombinant mutant S247M | Cylindrospermopsis raciborskii |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 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]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 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
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
recombinant mutant F245N/S247M | Cylindrospermopsis raciborskii |
| 8 | - |
recombinant mutants F245N and S247M | Cylindrospermopsis raciborskii |
| 8.5 | - |
recombinant wild-type CyrA | Cylindrospermopsis raciborskii |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 7 | 9 | above pH 9, the activity dropped rapidly for all enzymes | Cylindrospermopsis raciborskii |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | ligand-induced conformational changes in wild-type CyrA, structure-function-stability relationship, overview | Cylindrospermopsis raciborskii |
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