Enzymatic diagnosis of 3-hydroxy-3-methylglutaryl-CoA lyase deficiency with high-performance liquid chromatography.
A new non-radiochemical method for determination of 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) lyase is described. Acetyl-CoA, the product of the enzymatic reaction, is separated from the substrate by high-performance liquid chromatography and is quantified. The mean 3-hydroxy-3-methylglutaryl-CoA lyase activity in control fibroblasts was 7.8 +/- 2.1 (SD) nmol/min per mg protein, and its apparent Km value was 77.8 +/- 14.3 microM (R/S mixture) with a calculated Vmax of 12.4 +/- 2.2 nmol/min per mg protein. Using this method, we could easily differentiate a patient with 3-hydroxy-3-methylglutaryl-CoA lyase deficiency from control subjects.
Kikuchi
M
M
Department of Pediatrics, Tohoku University School of Medicine, Sendai, Japan.
Narisawa
K
K
Tada
K
K
Sweetman
L
L
D000105:Acetyl Coenzyme A / Q000378:metabolism; D000214:Acyl Coenzyme A / Q000378:metabolism; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D007223:Infant; D007700:Kinetics; D008661:Metabolism, Inborn Errors / Q000175:diagnosis; D007652:Oxo-Acid-Lyases / Q000172:deficiency*
D000214:Acyl Coenzyme A; C008047:3-hydroxy-3-methylglutaryl-coenzyme A; D000105:Acetyl Coenzyme A; D007652:Oxo-Acid-Lyases; C021410:3-hydroxy-3-methylglutaryl-coenzyme A lyase