Allosteric activation by nucleotides of the inactive by phosphatase ornithine decarboxylase of Escherichia coli.
ODC was purified to homogeneity from E. coli K12 MG1655 strain transformed with a pBR322 plasmid carrying the ODC gene. This preparation was homogeneous as it was analyzed by SDS-polyacrylamide gel electrophoresis. From this preparation the amino-terminal sequence analysis was obtained. The native ODC of E. coli is activated by ATP, GTP, CTP and UTP at 10(-3) M concentration to around 170-300%. Our results indicate that the recombinant ODC is activated only by GTP and UTP at 10(-3) M 370% and 300%, respectively. When the recombinant ODC was incubated with calf intestine alkaline phosphatase, this inactive ODC can be reversibly activated allosterically only by GTP or UTP at a concentration of 10(-6) or 10(-5) M. That GTP or UTP can allosterically convert the inactive form of ODC to an active form suggests that these analogues may be the in vivo physiological regulators of ODC.
Anagnostopoulos
C
C
Laboratory of Biochemistry, Faculty of Chemistry, Aristotelian University of Thessaloniki, Greece.
Choli
T
T
Kyriakidis
D A
DA
D000469:Alkaline Phosphatase / Q000378:metabolism; D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology*; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000494:pharmacology*; D009955:Ornithine Decarboxylase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D065108:Ornithine Decarboxylase Inhibitors; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D009711:Nucleotides; D065108:Ornithine Decarboxylase Inhibitors; D011994:Recombinant Proteins; D000469:Alkaline Phosphatase; D009955:Ornithine Decarboxylase