Assay of methylmalonyl CoA mutase with high-performance liquid chromatography.
An assay for methylmalonyl CoA mutase activity is described. Succinyl CoA produced in this method is separated from the substrate, methylmalonyl CoA, by reverse-phase high-performance liquid chromatography and is quantified. This method is useful to differentiate mutase apoenzyme deficiency (mut0, mut-) and the defect in deoxyadenosylcobalamin synthesis using fibroblasts cultured in high concentration of supplementary hydroxocobalamin. In methylmalonic acidemia, measurement of lymphocytes mutase activity offers therapeutical and prognostic informations.
Kikuchi
M
M
Department of Pediatrics, Tohoku University School of Medicine, Sendai, Japan.
Hanamizu
H
H
Narisawa
K
K
Tada
K
K
D000214:Acyl Coenzyme A / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D007535:Isomerases / Q000302:isolation & purification*; D008214:Lymphocytes / Q000201:enzymology; D008765:Methylmalonyl-CoA Mutase / Q000302:isolation & purification* / Q000378:metabolism
D000214:Acyl Coenzyme A; C012046:succinyl-coenzyme A; D007535:Isomerases; D008765:Methylmalonyl-CoA Mutase