Literature summary extracted from
Takahashi, R.; Nakamura, S.; Yoshida, T.; Kobayashi, Y.; Ohkubo, T.
Crystallization of human nicotinamide phosphoribosyltransferase (2007), Acta Crystallogr. Sect. F, 63, 375-377.
Application
EC Number |
Application |
Comment |
Organism |
---|
2.4.2.12 |
drug development |
inhibitor-soaking experiments |
Homo sapiens |
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
2.4.2.12 |
cDNA into pGEX6p1 for expression in Escherichia coli BL21 (DE) as glutathione S-transferase (GST) tagged fusion protein |
Homo sapiens |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
2.4.2.12 |
apo enzyme, diffraction of third crystals at 2 A resolution at -174°C, crystals belong to space group P2(1) with unit-cell parameters a = 60.56 A, b = 106.40 A, c =82.78 A, beta = 96.50°, technique: 1 microlitre of 10 mg/ml protein solution to 1 microlitre reservoir solution (38% pentaerytritol propoxylate (5/4 PO/OH) and 200 mM KCl pH 9) one week at 4°C, generation of first crystals by hanging-drop vapour diffusion (Index HT screening kit), generation of second crystals by microseeding: reservoir solution contained crushed first crystals, generation of third crystals by Repeated Seeding Technique: reservoir solution contained crushed second crystals |
Homo sapiens |
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
2.4.2.12 |
nicotinamide + alpha-D-5-phosphoribosyl-1-diphosphate |
Homo sapiens |
- |
nicotinamide mononucleotide + diphosphate |
- |
r |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
2.4.2.12 |
Homo sapiens |
P43490 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
2.4.2.12 |
lysis of bacteria by ultrasonification in presence of 1 mM phenylmethylsulfonylfluorid (PMSF) and 1 mM dithiothreitol (DTT), ultracentrifugation, supernatant applied to glutathione Sepharose 4B beads and washed, cleavage of glutathione S-transferase (GST) tag by PreScission Protease, purification by gel-filtration chromatography in Tris-HCl pH 8.5, 150 mM NaCl, 1 mM DTT, concentration to 10 mg/ml by ultrafiltration on Microcon YM-10, stored at 4°C |
Homo sapiens |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
2.4.2.12 |
nicotinamide + alpha-D-5-phosphoribosyl-1-diphosphate |
- |
Homo sapiens |
nicotinamide mononucleotide + diphosphate |
- |
r |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
2.4.2.12 |
dimer |
according to sedimentation-equilibrium measurements following ultracentrifugation of the protein solution |
Homo sapiens |
2.4.2.12 |
homodimer |
two molecules in the asymmetric unit of the crystal according to Matthews coefficient Vm = 2.4 A/Da, corresponding to a solvent content of 48.5% |
Homo sapiens |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
2.4.2.12 |
nicotinamide phosphoribosyltransferase |
- |
Homo sapiens |
2.4.2.12 |
NMPRTase |
- |
Homo sapiens |