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Literature summary for 1.1.1.302 extracted from

  • Chatwell, L.; Krojer, T.; Fidler, A.; Römisch, W.; Eisenreich, W.; Bacher, A.; Huber, R.; Fischer, M.J.
    Biosynthesis of riboflavin: structure and properties of 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5'-phosphate reductase of Methanocaldococcus jannaschii (2006), Mol. Biol., 359, 1334-1351.
    View publication on PubMed

Cloned(Commentary)

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 (Commentary)

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 [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

Organism UniProt Comment Textmining
Methanocaldococcus jannaschii
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant protein Methanocaldococcus jannaschii

Reaction

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 [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0008
-
-
Methanocaldococcus jannaschii

Substrates and Products (Substrate)

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

Subunits Comment Organism
homodimer 2 * 24906, electrospray mass spectrometry Methanocaldococcus jannaschii
homodimer 2 * 25037, calculated mass of the full-length protein Methanocaldococcus jannaschii

Synonyms

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Methanocaldococcus jannaschii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Methanocaldococcus jannaschii

Cofactor

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