Purine nucleotide pyrophosphotransferase from Streptomyces morookaensis, capable of synthesizing pppApp and pppGpp.
Purine nucleotide pyrophosphotransferase was purified to apparent homogeneity from a culture filtrate of Streptomyces morookaensis. It is a monomeric protein with a molecular weight of 24 000-25 000, and its isoelectric point is 6.9. The enzyme synthesizes purine nucleoside 5'-phosphate (mono, di, or tri) 3'-diphosphates such as pppApp, ppApp, pApp, pppGpp, ppGpp and pppIpp by transferring a pyrophosphoryl group from the 5'-position of ATP, dATP and ppApp to the 3'-position of purine nucleotides. The purified enzyme catalysed the formation of 435 mumol of pppApp and 620 mumol of pppGpp from ATP and GTP per min mg protein under the standard conditions. The enzyme requires absolutely a divalent cation for activity, and optimum pH for the enzyme activity lay above 10 for Mg2+, for Co2+ and Zn2+ from 9 to 9.5, and for Fe2+ from 7.5 to 8. The following Michaelis constants were determined: AMP, 2.78 mM; ADP, 3.23 mM; GMP, 0.89 mM; GDP, 0.46 mM and GTP, 1.54 mM, in the case of ATP donor. The enzyme is inhibited by guanine, guanosine, dGDP, dGTP, N-bromosuccinimide, iodacetate, sodium borate and mercuric acetate.
D000255:Adenosine Triphosphate / Q000378:metabolism; D001881:Borates / Q000494:pharmacology; D001974:Bromosuccinimide / Q000494:pharmacology; D002413:Cations, Divalent; D011756:Diphosphates / Q000378:metabolism; D017854:Diphosphotransferases; D006147:Guanine / Q000494:pharmacology; D006151:Guanosine / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007461:Iodoacetates / Q000494:pharmacology; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D010770:Phosphotransferases / Q000302:isolation & purification / Q000378:metabolism*; D011685:Purine Nucleotides / Q000096:biosynthesis*; D013302:Streptomyces / Q000201:enzymology*
D001881:Borates; D002413:Cations, Divalent; D011756:Diphosphates; D007461:Iodoacetates; D011685:Purine Nucleotides; D006151:Guanosine; D006147:Guanine; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D017854:Diphosphotransferases; C022482:purine nucleotide pyrophosphotransferase; D008274:Magnesium; D001974:Bromosuccinimide