Tandem mass spectrometric assay for the determination of carnitine palmitoyltransferase II activity in muscle tissue
- PMID: 11878804
- DOI: 10.1006/abio.2001.5554
Tandem mass spectrometric assay for the determination of carnitine palmitoyltransferase II activity in muscle tissue
Abstract
Carnitine palmitoyltransferase II (CPT-II) mediates the import of long-chain fatty acids into the mitochondrial matrix for subsequent beta-oxidation. Defects of CPT-II manifest as a severe neonatal hepatocardiomuscular form or as a mild muscular phenotype in early infancy or adolescence. CPT-II deficiency is diagnosed by the determination of enzyme activity in tissues involving the time-dependent conversion of radiolabeled CPT-II substrates (isotope-exchange assays) or the formation of chromogenic reaction products. We have established a mass spectrometric assay (MS/MS) for the determination of CPT-II activity based on the stoichiometric formation of acetylcarnitine in a coupled reaction system. In this single-tube reaction system palmitoylcarnitine is converted by CPT-II to free carnitine, which is subsequently esterified to acetylcarnitine by carnitine acetyltransferase. The formation of acetylcarnitine directly correlates with the CPT-II activity. Comparison of the MS/MS method (y) with our routine spectrophotometric assay (x) revealed a linear regression of y = 0.58x + 0.12 (r = 0.8369). Both assays allow one to unambiguously detect patients with the muscular form of CPT-II deficiency. However, the higher specificity and sensitivity as well as the avoidance of the drawbacks inherent in the use of radiolabeled substrates make this mass spectrometric method most suitable for the determination of CPT-II activity.
(C)2002 Elsevier Science (USA).
Similar articles
-
Carnitine palmitoyltransferase I activity monitoring in fibroblasts and leukocytes using electrospray ionization mass spectrometry.Anal Biochem. 1998 Feb 15;256(2):178-84. doi: 10.1006/abio.1997.2510. Anal Biochem. 1998. PMID: 9473275
-
[Heterogeneity of carnitine palmitoyltransferase deficiencies. Deficiency of CPT I in the hepatic form and CPT II in the muscular form].Arch Fr Pediatr. 1985 Aug-Sep;42 Suppl 1:613-7. Arch Fr Pediatr. 1985. PMID: 4083994 French.
-
[Molecular analysis of a patient with carnitine palmitoyltransferase II deficiency].Rinsho Shinkeigaku. 1997 Jun;37(6):532-5. Rinsho Shinkeigaku. 1997. PMID: 9366186 Japanese.
-
Muscle Carnitine Palmitoyltransferase II Deficiency: A Review of Enzymatic Controversy and Clinical Features.Int J Mol Sci. 2017 Jan 3;18(1):82. doi: 10.3390/ijms18010082. Int J Mol Sci. 2017. PMID: 28054946 Free PMC article. Review.
-
Muscle Carnitine Palmitoyltransferase II (CPT II) Deficiency: A Conceptual Approach.Molecules. 2020 Apr 13;25(8):1784. doi: 10.3390/molecules25081784. Molecules. 2020. PMID: 32295037 Free PMC article. Review.
Cited by
-
The enzymology of mitochondrial fatty acid beta-oxidation and its application to follow-up analysis of positive neonatal screening results.J Inherit Metab Dis. 2010 Oct;33(5):479-94. doi: 10.1007/s10545-010-9104-8. Epub 2010 May 20. J Inherit Metab Dis. 2010. PMID: 20490924 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
