Literature summary for 1.1.1.302 extracted from

Chatwell, L.; Krojer, T.; Fidler, A.; R�misch, 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.

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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

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	a

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