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2.4.2.12: nicotinamide phosphoribosyltransferase

This is an abbreviated version!
For detailed information about nicotinamide phosphoribosyltransferase, go to the full flat file.

Word Map on EC 2.4.2.12

Reaction

nicotinamide D-ribonucleotide
+
diphosphate
=
nicotinamide
 +
5-phospho-alpha-D-ribose 1-diphosphate

Synonyms

eNAMPT, extracellular nicotinamide phosphoribosyltrasferase, iNAMPT, Nam phosphoribosyltransferase, NAmPRTase, Nampt, Nampt/PBEF/Visfatin, Nampt/visfatin, NAMPTA, NAMPTB, NAPRT, nicotinamide mononucleotide pyrophosphorylase, nicotinamide mononucleotide synthetase, nicotinamide phosphoribosyl transferase, nicotinamide phosphoribosyltransferase, nicotinamide phosphoribosyltransferase/Visfatin, NMN pyrophosphorylase, NMN synthetase, NMPRTase, PBEF, phosphoribosyltransferase, nicotinamide, pre-B cell colony enhancing factor, pre-B cell colony enhancing factor(PBEF), pre-B cell colony-enhancing factor, pre-B cell colony-enhancing factor (PBEF), pre-B cell colony-enhancing factor 1, pre-B colony enhancing factor, pre-B-cell colony enhancing factor, pre-B-cell colony-enhancing factor, Visfatin

ECTree

     2 Transferases
         2.4 Glycosyltransferases
             2.4.2 Pentosyltransferases
                2.4.2.12 nicotinamide phosphoribosyltransferase

Purification

Purification on EC 2.4.2.12 - nicotinamide phosphoribosyltransferase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
immobilized metal ion affinity chromatography
-
immobilized metal ion affinity chromatography (Ni2+)
lysis of bacteria by French press, elution from Ni-NTA column by 100 and 200 mM imidazole, concentration and desalting, elution from HiLoad Superdex 200GP 26/60 column with 100 mM HEPES pH 7.5, 100 mM NaCl, 10 mM beta-mercaptoethanol, concentrated to 50-75 mg/ml, kept frozen
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
Ni-NTA resin column chromatography
partial
partially frim K-562 leukemic cells
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography