Regulation of the Escherichia coli biosynthetic ornithine decarboxylase activity by phosphorylation and nucleotides.
A protein kinase which phosphorylates in vitro the biosynthetic ornithine decarboxylase (ODC) was partially purified from Escherichia coli. In vivo phosphorylation of ODC occurs after incubation of E. coli with [32P]orthophosphate. When the recombinant ODC was incubated with calf intestine alkaline phosphatase it was inactivated and this inactive ODC could be reversibly activated allosterically only by guanyl or uracyl phosphate analogues at a concentration of 10(-4) or 10(-3) M. The pH optimum of the [8-3H]GTP binding was determined and it was shown that the GTP binding is proportional to the amount of ODC. The [8-3H]GTP binds specifically to ODC as was proved by the addition of cold GTP or ATP. High concentration of GTP can dissociate the ODC-antizyme complex and either reactivate or liberate the ODC. Therefore, a working hypothesis is suggested describing the regulation of ODC by phosphorylation-dephosphorylation reaction or by antizyme and nucleotide analogues interaction.
Anagnostopoulos
C G
CG
Department of Chemistry, Aristotle University of Thessaloniki, Greece.
Kyriakidis
D A
DA
D002852:Chromatography, Ion Exchange; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D004926:Escherichia coli / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D009711:Nucleotides / Q000494:pharmacology*; D009955:Ornithine Decarboxylase / Q000302:isolation & purification / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D011233:Precipitin Tests; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic / Q000235:genetics
D009711:Nucleotides; D011994:Recombinant Proteins; D011494:Protein Kinases; D010744:Phosphoric Monoester Hydrolases; D009955:Ornithine Decarboxylase