Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant enzymes from plasmids in strain DH5alpha | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
C110G | site-directed mutagenesis, 0.22% activity compared to the wild-type enzyme, increased temperature optimum | Escherichia coli |
C181S | site-directed mutagenesis, 0.29% activity compared to the wild-type enzyme, highly increased temperature optimum | Escherichia coli |
E111K | site-directed mutagenesis, 3.1% activity compared to the wild-type enzyme, increased temperature optimum | Escherichia coli |
E152K | site-directed mutagenesis, 0.06% activity compared to the wild-type enzyme, increased pH optimum, decreased temperature optimum | Escherichia coli |
H112D | site-directed mutagenesis, 0.23% activity compared to the wild-type enzyme, decreased pH optimum, increased temperature optimum | Escherichia coli |
H113N | site-directed mutagenesis, 67% activity compared to the wild-type enzyme, decreased pH optimum, increased temperature optimum | Escherichia coli |
H179Q | site-directed mutagenesis, 0.8% activity compared to the wild-type enzyme, increased temperature optimum | Escherichia coli |
K136E | site-directed mutagenesis, 0.25% activity compared to the wild-type enzyme, increased temperature optimum | Escherichia coli |
L134Q | site-directed mutagenesis, 1.85% activity compared to the wild-type enzyme, increased temperature optimum | Escherichia coli |
R139C | site-directed mutagenesis, 29.3% activity compared to the wild-type enzyme, decreased pH optimum | Escherichia coli |
R185G | site-directed mutagenesis, 2.9% activity compared to the wild-type enzyme, decreased temperature optimum | Escherichia coli |
R56L | site-directed mutagenesis, 14% activity compared to the wild-type enzyme, increased temperature optimum | Escherichia coli |
S135C | site-directed mutagenesis, 0.7% activity compared to the wild-type enzyme, decreased pH and temperature optimum | Escherichia coli |
V150E | site-directed mutagenesis, 3.2% activity compared to the wild-type enzyme, slightly decreased pH optimum, increased temperature optimum | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | association and dissociation constants of wild-type and mutant enzymes in oligomerization | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
additional information | - |
MW of mutant E152K is very high in gel filtration approximately corresponding to a didecameric form | Escherichia coli |
25000 | - |
10 * 25000, about, composed of a pentamers of 5 dimers, the active site is located at the interface between dimers, Lys136, Arg139, and Glu152 are especially important for the oligomerization | Escherichia coli |
50000 | - |
dimeric mutants K136E and R139C, gel filtration | Escherichia coli |
120000 | - |
wild-type enzyme in presence of 0.3 M KCl, mutants C110G, E111K, H112D, and C181S, gel filtration | Escherichia coli |
250000 | - |
decameric wild-type enzyme, gel filtration | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + H2O | Escherichia coli | first step in the biosynthesis of pteridine coenzymes, such as folic acid and tetrahydrobiopterin | dihydroneopterin triphosphate + formate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes | Escherichia coli |
Renatured (Comment) | Organism |
---|---|
reconstitution of dissociated mutant enzyme subunits to chimeric dimers from 2 monomers A and B derived from 2 different mutants, overview | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
reconstituted chimeric mutant dimers, overview | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + H2O | - |
Escherichia coli | dihydroneopterin triphosphate + formate | - |
? | |
GTP + H2O | first step in the biosynthesis of pteridine coenzymes, such as folic acid and tetrahydrobiopterin | Escherichia coli | dihydroneopterin triphosphate + formate | - |
? |
Subunits | Comment | Organism |
---|---|---|
decamer | 10 * 25000, about, composed of a pentamers of 5 dimers, the active site is located at the interface between dimers, Lys136, Arg139, and Glu152 are especially important for the oligomerization | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
GTP CHase I | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
mutant E152K | Escherichia coli |
55 | - |
mutants S135C and R185G | Escherichia coli |
60 | - |
wild-type enzyme and mutant R139C | Escherichia coli |
65 | - |
mutants H112D, K136E, and H179Q | Escherichia coli |
70 | - |
mutant H113 N | Escherichia coli |
72 | - |
mutant R56L | Escherichia coli |
75 | - |
mutants C110G and L134Q | Escherichia coli |
85 | - |
mutants E111K and V150E | Escherichia coli |
90 | - |
mutant C181S | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
mutant S135C | Escherichia coli |
7 | 8.5 | mutant V150E | Escherichia coli |
7.5 | - |
mutant H112D | Escherichia coli |
8 | - |
mutants H113N and R139C | Escherichia coli |
8.5 | - |
wild-type enzyme and mutants R56L, C110G, E111K, L134Q, K136E, H179Q, C181S, and R185G | Escherichia coli |
9 | - |
mutant E152K | Escherichia coli |