A radioisotopic method for fructose-1-phosphate aldolase assay that facilitates diagnosis of hereditary fructose intolerance.
A sensitive new method in which D-[U-14C]fructose-1-phosphate is used for fructose-bisphosphate aldolase (EC 2.1.2.13) assay is described. The radioactive fructose-1-phosphate compound was prepared from [U-14C]fructose by use of partly purified fructokinase (EC 2.7.1.4). With this method we measured normal values for aldolase in human liver (2.4-10.0 nmol/min per mg of protein), kidney (3.6-3.8), and intestine (4.2-10.0) as well as Km values for fructose-1-phosphate (approximately 1.0-2.2 mmol/L). In patients with hereditary fructose intolerance the aldolase activity in liver and intestine was less than 10% of normal values. The Lineweaver-Burk plots for data from patients with hereditary fructose intolerance were hyperbolic, indicating a structural alteration in the enzyme.
D000328:Adult; D002239:Carbohydrate Metabolism, Inborn Errors / Q000175:diagnosis*; D002250:Carbon Radioisotopes; D002648:Child; D004796:Clinical Enzyme Tests; D005260:Female; D005633:Fructose Intolerance / Q000175:diagnosis*; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis*; D006801:Humans; D007223:Infant; D007422:Intestines / Q000201:enzymology; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D013053:Spectrophotometry
D002250:Carbon Radioisotopes; D005634:Fructose-Bisphosphate Aldolase