AMP deaminase in Dictyostelium discoideum: increase in activity following nutrient deprivation induced by starvation or hadacidin.
AMP deaminase, the activity that catalyzes the deamination of AMP to form IMP and NH3 has been measured in Dictyostelium discoideum. A new procedure to assay the activity of this enzyme was developed using formycin 5'-monophosphate, a fluorescent analog of AMP as the substrate, and ion-paired reverse phase HPLC to separate the reactants and products. Quantitation of the formycin containing compounds was accomplished at 290 nm. At this wavelength adenosine containing compounds were not detected and activity could be monitored in the presence of its activator ATP. The AMP deaminase activity in vegetative cells was 7.4 nmols/min/mg proteins while the activity in cells measured at 2 and 6 hrs after starvation-induced growth-arrest was 376 nmols/min/mg protein...a 51-fold increase. When vegetative cells were treated with hadacidin, a drug that restricts de novo AMP synthesis and pinocytosis, the activity of the AMP deaminase was 511 nmols/min/mg protein...a 70-fold increase compared to that in untreated vegetative cells. Smaller increases were noted following the inhibition of growth with the drugs cerulenin and vinblastine, as well as after the inhibition of de novo GMP synthesis with the drug mycophenolic acid or the partial inhibition of de novo AMP synthesis with analogs of hadacidin, N-hydroxyglycine and N-formylglycine. In addition, when the activity of two other enzymes involved in purine metabolism, namely adenosine kinase and hypoxanthine-guanine phosphoribosyl transferase, was measured in vegetative cells, and the activity of both compared to that measured in starvation and hadacidin induced growth-arrested cells, showed no significant changes.(ABSTRACT TRUNCATED AT 250 WORDS)
D000659:AMP Deaminase / Q000378:metabolism*; D000248:Adenosine Kinase / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D003470:Culture Media; D004023:Dictyostelium / Q000201:enzymology* / Q000254:growth & development; D004789:Enzyme Activation; D005573:Formycins / Q000378:metabolism; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006157:Guanosine Monophosphate / Q000096:biosynthesis; D007041:Hypoxanthine Phosphoribosyltransferase / Q000378:metabolism; D009709:Nucleotide Deaminases / Q000378:metabolism*; D012265:Ribonucleotides / Q000378:metabolism; D013379:Substrate Specificity
D003470:Culture Media; D005573:Formycins; D012265:Ribonucleotides; C021683:formycin 5'-phosphate; D000249:Adenosine Monophosphate; D006157:Guanosine Monophosphate; D000255:Adenosine Triphosphate; C001321:hadacidin; D007041:Hypoxanthine Phosphoribosyltransferase; D000248:Adenosine Kinase; D009709:Nucleotide Deaminases; D000659:AMP Deaminase; D005998:Glycine