A radioisotopic assay for polyamine oxidase.
A new radioisotopic assay for polyamine oxidase with N1-acetylspermine as substrate is presented. A modified method for the chemical synthesis of radioactive N1-acetylspermine, which gave a good yield, is also described. The reaction mixture, containing N1-[14C]acetylspermine and tissue homogenate, was incubated for the enzyme reaction and applied to a minicolumn of Amberlite CG-50. The reaction product 3-[14C]-acetamidopropanal did not adsorb to the column, but passed through it; thus the eluate could be directly subjected to liquid scintillation counting. The blank levels were low and relatively constant even with crude tissue homogenates. The detection limit obtained was 0.05 nmol per tube. This method is simple, highly sensitive, and highly specific.
Kumazawa
T
T
Department of Legal Medicine, Hamamatsu University School of Medicine, Japan.
Seno
H
H
Suzuki
O
O
D000818:Animals; D002855:Chromatography, Thin Layer; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007553:Isotope Labeling; D008297:Male; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000032:analysis* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013050:Spectrometry, Fluorescence; D013095:Spermidine / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000378:metabolism; D000097667:Polyamine Oxidase
D000587:Oxidoreductases Acting on CH-NH Group Donors; D013095:Spermidine; D000097667:Polyamine Oxidase; C017988:N(1)-acetylspermidine