Cloned (Comment) | Organism |
---|---|
expression of wild-type and several mutants in Escherichia coli | Rattus norvegicus |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapour diffusion, polyethylene glycol 4500, Tris phosphate, pH 7.8, 10% methanol, 4°C, crystals of binary complexes with NADH of recombinant wild-type, W86I, Y146F, Y146H, Y146F/K150Q and A133S mutant enzymes, 2.1-2.3 A resolution | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
A133S | similar kcat for quinonoid dihydrobiopterin as wild-type | Rattus norvegicus |
D37I | lower kcat for quinonoid dihydrobiopterin than wild-type | Rattus norvegicus |
K150I | lower kcat for quinonoid dihydrobiopterin than wild-type | Rattus norvegicus |
K150M | lower kcat for quinonoid dihydrobiopterin than wild-type | Rattus norvegicus |
K150Q | lower kcat for quinonoid dihydrobiopterin than wild-type | Rattus norvegicus |
N186A | lower kcat for quinonoid dihydrobiopterin than wild-type | Rattus norvegicus |
W86I | lower kcat for quinonoid dihydrobiopterin than wild-type | Rattus norvegicus |
Y146F | almost no activity with quinonoid dihydrobipterin | Rattus norvegicus |
Y146F/K150Q | lower kcat for quinonoid dihydrobiopterin than wild-type | Rattus norvegicus |
Y146H | lower kcat for quinonoid dihydrobiopterin than wild-type | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0009 | - |
quinonoid dihydrobiopterin | Y146F mutant enzyme | Rattus norvegicus | |
0.01 | - |
NADH | Y146F mutant enzyme | Rattus norvegicus | |
0.012 | - |
NADH | W86I mutant enzyme | Rattus norvegicus | |
0.013 | - |
NADH | recombinant wild-type enzyme | Rattus norvegicus | |
0.017 | - |
NADH | Y146H mutant enzyme | Rattus norvegicus | |
0.02 | - |
quinonoid dihydrobiopterin | A133S mutant enzyme | Rattus norvegicus | |
0.021 | - |
NADH | K150I mutant enzyme | Rattus norvegicus | |
0.023 | - |
quinonoid dihydrobiopterin | Y146H mutant enzyme | Rattus norvegicus | |
0.025 | - |
NADH | A133S mutant enzyme | Rattus norvegicus | |
0.026 | - |
quinonoid dihydrobiopterin | K150I mutant enzyme | Rattus norvegicus | |
0.027 | - |
quinonoid dihydrobiopterin | recombinant wild-type enzyme | Rattus norvegicus | |
0.027 | - |
NADH | Y146F/K150Q double mutant enzyme | Rattus norvegicus | |
0.029 | - |
NADH | K150Q mutant enzyme | Rattus norvegicus | |
0.03 | - |
quinonoid dihydrobiopterin | D37I mutant enzyme | Rattus norvegicus | |
0.033 | - |
quinonoid dihydrobiopterin | K150Q mutant enzyme | Rattus norvegicus | |
0.036 | - |
NADH | D37I mutant enzyme | Rattus norvegicus | |
0.039 | - |
quinonoid dihydrobiopterin | K150M mutant enzyme | Rattus norvegicus | |
0.041 | - |
quinonoid dihydrobiopterin | Y146F/K150Q double mutant enzyme | Rattus norvegicus | |
0.07 | - |
quinonoid dihydrobiopterin | W86I mutant enzyme | Rattus norvegicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and several mutant enzymes | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
quinoid dihydrobiopterin + NAD(P)H | - |
Rattus norvegicus | tetrahydrobiopterin + NAD(P)+ | - |
? | |
quinonoid dihydrobiopterin + NADH + H+ | - |
Rattus norvegicus | 6R-(1'R,2'R)-5,6,7,8-tetrahydrobiopterin + NAD+ | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.7 | - |
quinoid dihydrobiopterin | Y146F mutant enzyme | Rattus norvegicus | |
23 | - |
quinoid dihydrobiopterin | Y146H mutant enzyme | Rattus norvegicus | |
26 | - |
quinoid dihydrobiopterin | K150M mutant enzyme | Rattus norvegicus | |
36 | - |
quinoid dihydrobiopterin | D37I mutant enzyme | Rattus norvegicus | |
64 | - |
quinoid dihydrobiopterin | W86I mutant enzyme | Rattus norvegicus | |
64 | - |
quinoid dihydrobiopterin | K150I mutant enzyme | Rattus norvegicus | |
76 | - |
quinoid dihydrobiopterin | Y146F/K150Q double mutant enzyme | Rattus norvegicus | |
90 | - |
quinoid dihydrobiopterin | K150Q mutant enzyme | Rattus norvegicus | |
149 | - |
quinoid dihydrobiopterin | A133S mutant enzyme | Rattus norvegicus | |
153 | - |
quinoid dihydrobiopterin | recombinant wild-type enzyme | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
recombinant wild-type, D37I, W86I, Y146F, Y146H, K150Q, K150I, K150M, N186A, A133S and Y146F/K150Q mutant enzymes | Rattus norvegicus |