Enzymatic diagnosis of 3-hydroxy-3-methylglutaryl-CoA lyase deficiency with high-performance liquid chromatography

Clin Chim Acta. 1990 Aug 31;189(3):297-301. doi: 10.1016/0009-8981(90)90311-f.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyl Coenzyme A / metabolism
  • Acyl Coenzyme A / metabolism
  • Child, Preschool
  • Chromatography, High Pressure Liquid / methods
  • Humans
  • Infant
  • Kinetics
  • Metabolism, Inborn Errors / diagnosis
  • Oxo-Acid-Lyases / deficiency*

Substances

  • Acyl Coenzyme A
  • 3-hydroxy-3-methylglutaryl-coenzyme A
  • Acetyl Coenzyme A
  • Oxo-Acid-Lyases
  • 3-hydroxy-3-methylglutaryl-coenzyme A lyase