Application | Comment | Organism |
---|---|---|
drug development | inhibitor-soaking experiments | Homo sapiens |
Cloned (Comment) | Organism |
---|---|
cDNA into pGEX6p1 for expression in Escherichia coli BL21 (DE) as glutathione S-transferase (GST) tagged fusion protein | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
nicotinamide + alpha-D-5-phosphoribosyl-1-diphosphate | Homo sapiens | - |
nicotinamide mononucleotide + diphosphate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P43490 | - |
- |
Purification (Comment) | Organism |
---|---|
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
nicotinamide + alpha-D-5-phosphoribosyl-1-diphosphate | - |
Homo sapiens | nicotinamide mononucleotide + diphosphate | - |
r |
Subunits | Comment | Organism |
---|---|---|
dimer | according to sedimentation-equilibrium measurements following ultracentrifugation of the protein solution | Homo sapiens |
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 | Comment | Organism |
---|---|---|
nicotinamide phosphoribosyltransferase | - |
Homo sapiens |
NMPRTase | - |
Homo sapiens |