Assays for amino acid decarboxylase enzymes using ion-exchange cartridges.
A general radiochemical method for estimating the activity of amino acid decarboxylases is reported. This method utilizes ion-exchange cartridges to separate unreacted radiolabeled amino acid substrates from product amines, which can then readily be quantitated by liquid scintillation counting. The assay is simple, rapid, and more sensitive than standard 14CO2 trapping procedures if uniformly labeled amino acid substrates are utilized. Acidic, basic, and aromatic amino acid decarboxylases can be assayed with the appropriate choice of cation or anion exchangers. The utility of the method is demonstrated for aspartate-alpha-decarboxylase, tyrosine decarboxylase, and lysine decarboxylase where kinetic parameters are comparable to values obtained by standard radiochemical 14CO2 trapping assays.
Heerze
L D
LD
Department of Food Science, University of Alberta, Edmonton, Canada.
Kang
Y J
YJ
Palcic
M M
MM
D000596:Amino Acids / Q000032:analysis; D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000032:analysis; D002262:Carboxy-Lyases / Q000032:analysis*; D002852:Chromatography, Ion Exchange / Q000379:methods; D005968:Glutamate Decarboxylase / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D008722:Methods; D014445:Tyrosine Decarboxylase / Q000032:analysis*
D000596:Amino Acids; D002262:Carboxy-Lyases; C019104:aspartate-alpha-decarboxylase; D005968:Glutamate Decarboxylase; C021520:lysine decarboxylase; D014445:Tyrosine Decarboxylase; D001142:Aromatic-L-Amino-Acid Decarboxylases