Nucleotide phosphotransferase of Escherichia coli: purification by affinity chromatography.
Improved extraction and purification procedures permit the isolation from Escherichia coli W cells of much larger quantities and of more highly purified preparations of nucleotide phosphotransferase. Of various affinity resins tested for efficiency of purification, columns of agarose/5'-AMP (AGAMP), type 3, proved the best. In this way a 300- to 450-fold purification of the enzyme was achieved in a few steps. The enzyme, which, as reported before, transfers organically bound phosphate to the 2' or 3' hydroxyls of nucleosides and nucleotides, was tested in its behavior toward a series of ribonucleosidonucleotides, namely, CpC, ApA, CpA, and ApC. All were phosphate acceptors, but a detailed comparative study of adenosine and cytidine, 5'-AMP and 5'-CMP, and ApA and ApC revealed peculiar specificities in the relative distribution of the phosphorylated products.
D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000201:enzymology*; D009243:NAD; D010770:Phosphotransferases / Q000302:isolation & purification* / Q000378:metabolism; D012263:Ribonucleosides; D012265:Ribonucleotides; D013379:Substrate Specificity
D012263:Ribonucleosides; D012265:Ribonucleotides; D009243:NAD; D010770:Phosphotransferases; C021898:nucleotide phosphotransferase