Method for Measurement of Peroxisomal Very Long-Chain Fatty Acid Beta-Oxidation and De Novo C26:0 Synthesis Activity in Living Cells Using Stable-Isotope Labeled Docosanoic Acid

Malu-Clair van de Beek; Inge M E Dijkstra; Stephan Kemp

doi:10.1007/978-1-4939-6937-1_5

Method for Measurement of Peroxisomal Very Long-Chain Fatty Acid Beta-Oxidation and De Novo C26:0 Synthesis Activity in Living Cells Using Stable-Isotope Labeled Docosanoic Acid

Methods Mol Biol. 2017:1595:45-54. doi: 10.1007/978-1-4939-6937-1_5.

Authors

Malu-Clair van de Beek¹, Inge M E Dijkstra¹, Stephan Kemp²

Affiliations

¹ Laboratory Genetic Metabolic Diseases (F0-226), Departments of Pediatrics and Clinical Chemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands.
² Laboratory Genetic Metabolic Diseases (F0-226), Departments of Pediatrics and Clinical Chemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands. [email protected].

PMID: 28409450
DOI: 10.1007/978-1-4939-6937-1_5

Abstract

Peroxisomes are present in virtually every eukaryotic cell type with the exception of the mature erythrocyte. In higher eukaryotes, one of the main functions of peroxisomes is lipid metabolism by means of beta-oxidation of very long-chain fatty acids (VLCFA; ≥22 carbon atoms). A dysfunction in peroxisomal VLCFA beta-oxidation results in elevated VLCFA levels in cells, tissue, and plasma. Here, we describe a straightforward and sensitive method to measure peroxisomal beta-oxidation capacity in living cells using stable-isotope labeled docosanoic acid (D₃-C22:0).

Keywords: Beta-oxidation; Elongation; Fatty acids; Peroxisome; Stable-isotope; Very-long-chain fatty acids.

MeSH terms

Cell Culture Techniques
Cells, Cultured
Fatty Acids / metabolism*
Fibroblasts / metabolism
Humans
Isotopes*
Lipid Metabolism*
Oxidation-Reduction*
Peroxisomes / metabolism*

Substances

Fatty Acids
Isotopes
behenic acid