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

Masahiro Kikuchi; Kuniaki Narisawa; Keiya Tada; Laurence Sweetman

doi:10.1016/0009-8981(90)90311-F

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

Masahiro Kikuchi, Kuniaki Narisawa, Keiya Tada, Laurence Sweetman

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

A new non-radiochemical method for determination of 3-hydroxy-3-methylglutaryl-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 K_m value was 77.8 ± 14.3 μM (R/S mixture) with a calculated V_max 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.

Original language	English (US)
Pages (from-to)	297-301
Number of pages	5
Journal	Clinica Chimica Acta
Volume	189
Issue number	3
DOIs	https://doi.org/10.1016/0009-8981(90)90311-F
State	Published - Aug 31 1990
Externally published	Yes

Keywords

3-Hydroxy-3-methylglutaric aciduria
High-performance liquid chromatography
Hydroxymethylglutaryl-CoA lyase

ASJC Scopus subject areas

Biochemistry
Clinical Biochemistry
Biochemistry, medical

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10.1016/0009-8981(90)90311-F

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title = "Enzymatic diagnosis of 3-hydroxy-3-methylglutaryl-CoA lyase deficiency with high-performance liquid chromatography",

abstract = "A new non-radiochemical method for determination of 3-hydroxy-3-methylglutaryl-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 μM (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.",

keywords = "3-Hydroxy-3-methylglutaric aciduria, High-performance liquid chromatography, Hydroxymethylglutaryl-CoA lyase",

author = "Masahiro Kikuchi and Kuniaki Narisawa and Keiya Tada and Laurence Sweetman",

note = "Funding Information: This work was supported by grants from the Ministry of Education, Culture and the Ministry of Health and Public Welfare, Japan.",

year = "1990",

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doi = "10.1016/0009-8981(90)90311-F",

language = "English (US)",

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T1 - Enzymatic diagnosis of 3-hydroxy-3-methylglutaryl-CoA lyase deficiency with high-performance liquid chromatography

AU - Kikuchi, Masahiro

AU - Narisawa, Kuniaki

AU - Tada, Keiya

AU - Sweetman, Laurence

N1 - Funding Information: This work was supported by grants from the Ministry of Education, Culture and the Ministry of Health and Public Welfare, Japan.

PY - 1990/8/31

Y1 - 1990/8/31

N2 - A new non-radiochemical method for determination of 3-hydroxy-3-methylglutaryl-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 μM (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.

AB - A new non-radiochemical method for determination of 3-hydroxy-3-methylglutaryl-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 μM (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.

KW - 3-Hydroxy-3-methylglutaric aciduria

KW - High-performance liquid chromatography

KW - Hydroxymethylglutaryl-CoA lyase

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JO - Clinica Chimica Acta

JF - Clinica Chimica Acta

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ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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