Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli using a synthetic gene, the synthetic open reading frame is optimized for expression in Escherichia coli. The hypothetical open reading frame MJ0671 of Methanocaldococcus jannaschii predicts a protein of 224 amino acid residues. Because MJ0671 contains numerous codons that are poorly translated in Escherichia coli, a synthetic gene is designed that is optimized for the Escherichia coli codon usage. Approximately 31% (69 of 224) of the codons are replaced, and 14 singular restriction sites are introduced. The DNA sequence is assembled from 16 synthetic oligonucleotides by a sequence of eight PCR steps. The synthetic gene is transcribed efficiently in a recombinant Escherichia coli strain, affording approximately 30% of cellular protein | Methanocaldococcus jannaschii |
Crystallization (Comment) | Organism |
---|---|
sitting-drop vapour diffusion method. The structure of the enzyme in complex with the cofactor nicotinamide adenine dinucleotide phosphate is determined by X-ray crystallography at a resolution of 2.5 A | Methanocaldococcus jannaschii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
24906 | - |
2 * 24906, electrospray mass spectrometry | Methanocaldococcus jannaschii |
25037 | - |
2 * 25037, calculated mass of the full-length protein | Methanocaldococcus jannaschii |
50000 | - |
analytical ultracentrifugation, boundary sedimentation | Methanocaldococcus jannaschii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanocaldococcus jannaschii | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant protein | Methanocaldococcus jannaschii |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2,5-diamino-6-(5-phospho-D-ribitylamino)pyrimidin-4(3H)-one + NAD(P)+ = 2,5-diamino-6-(5-phospho-D-ribosylamino)pyrimidin-4(3H)-one + NAD(P)H + H+ | the reaction proceeds in the opposite direction | Methanocaldococcus jannaschii |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0008 | - |
- |
Methanocaldococcus jannaschii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,5-diamino-6-(1-D-ribosylamino)-4(3H)-pyrimidinone 5'-phosphate + NADPH + H+ | the substrate, 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5'-phosphate, is modeled into the putative active site. The model suggests the transfer of the pro-R hydrogen of C-4 of NADPH to C-1' of the substrate | Methanocaldococcus jannaschii | 2,5-diamino-6-(1-D-ribitylamino)-4(3H)-pyrimidinone 5'-phosphate + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 24906, electrospray mass spectrometry | Methanocaldococcus jannaschii |
homodimer | 2 * 25037, calculated mass of the full-length protein | Methanocaldococcus jannaschii |
Synonyms | Comment | Organism |
---|---|---|
2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5'-phosphate reductase | - |
Methanocaldococcus jannaschii |
MJ0671 | - |
Methanocaldococcus jannaschii |
MjaRED | - |
Methanocaldococcus jannaschii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Methanocaldococcus jannaschii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Methanocaldococcus jannaschii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | the substrate, 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5'-phosphate, is modeled into the putative active site. The model suggests the transfer of the pro-R hydrogen of C-4 of NADPH to C-1' of the substrate | Methanocaldococcus jannaschii |