A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl-CoA hydratase deficiency
D-bifunctional protein (D-BP) plays an indispensable role in peroxisomal β-oxidation, and its inherited deficiency in humans is associated with severe clinical abnormalities. Three different subtypes of D-BP deficiency can be distinguished: 1) a complete deficiency of D-BP (type I), 2) an isolated D...
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doaj-46c44f7e4d09494884b0544422094e522021-04-27T04:39:18ZengElsevierJournal of Lipid Research0022-22752003-03-01443640644A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl-CoA hydratase deficiencyJolein Gloerich0Simone Denis1Elisabeth G. van Grunsven2Georges Dacremont3Ronald J.A. Wanders4Sacha Ferdinandusse5University of Amsterdam, Academic Medical Center, Departments of Clinical Chemistry and Pediatrics, Laboratory for Genetic Metabolic Diseases (F0-224), P.O. Box 22700, 1100 DE Amsterdam, The Netherlands; Department of Pediatrics, University of Ghent, Ghent, BelgiumUniversity of Amsterdam, Academic Medical Center, Departments of Clinical Chemistry and Pediatrics, Laboratory for Genetic Metabolic Diseases (F0-224), P.O. Box 22700, 1100 DE Amsterdam, The Netherlands; Department of Pediatrics, University of Ghent, Ghent, BelgiumUniversity of Amsterdam, Academic Medical Center, Departments of Clinical Chemistry and Pediatrics, Laboratory for Genetic Metabolic Diseases (F0-224), P.O. Box 22700, 1100 DE Amsterdam, The Netherlands; Department of Pediatrics, University of Ghent, Ghent, BelgiumUniversity of Amsterdam, Academic Medical Center, Departments of Clinical Chemistry and Pediatrics, Laboratory for Genetic Metabolic Diseases (F0-224), P.O. Box 22700, 1100 DE Amsterdam, The Netherlands; Department of Pediatrics, University of Ghent, Ghent, BelgiumUniversity of Amsterdam, Academic Medical Center, Departments of Clinical Chemistry and Pediatrics, Laboratory for Genetic Metabolic Diseases (F0-224), P.O. Box 22700, 1100 DE Amsterdam, The Netherlands; Department of Pediatrics, University of Ghent, Ghent, BelgiumUniversity of Amsterdam, Academic Medical Center, Departments of Clinical Chemistry and Pediatrics, Laboratory for Genetic Metabolic Diseases (F0-224), P.O. Box 22700, 1100 DE Amsterdam, The Netherlands; Department of Pediatrics, University of Ghent, Ghent, BelgiumD-bifunctional protein (D-BP) plays an indispensable role in peroxisomal β-oxidation, and its inherited deficiency in humans is associated with severe clinical abnormalities. Three different subtypes of D-BP deficiency can be distinguished: 1) a complete deficiency of D-BP (type I), 2) an isolated D-BP enoyl-CoA hydratase deficiency (type II), and 3) an isolated D-BP 3-hydroxyacyl-CoA dehydrogenase deficiency (type III). In this study, we developed a method to measure D-BP dehydrogenase activity independent of D-BP hydratase (D-BP HY) activity to distinguish between D-BP deficiency type I and type II, which until now was only possible by mutation analysis. For this assay, the hydratase domain of D-BP was expressed in the yeast Saccharomyces cerevisiae. After a coincubation of yeast homogenate expressing D-BP HY with fibroblast homogenate of patients using the enoyl-CoA ester of the bile acid intermediate trihydroxycholestanoic acid as substrate, D-BP dehydrogenase activity was measured. Fibroblasts of patients with a D-BP deficiency type II displayed D-BP dehydrogenase activity, whereas type I and type III patients did not.This newly developed assay to measure D-BP dehydrogenase activity in fibroblast homogenates provides a quick and reliable method to assign patients with deficient D-BP HY activity to the D-BP deficiency subgroups type I or type II.http://www.sciencedirect.com/science/article/pii/S0022227520312050peroxisomal fatty acid oxidation disordersvery long chain fatty acidstrihydroxycholestanoic acid |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jolein Gloerich Simone Denis Elisabeth G. van Grunsven Georges Dacremont Ronald J.A. Wanders Sacha Ferdinandusse |
spellingShingle |
Jolein Gloerich Simone Denis Elisabeth G. van Grunsven Georges Dacremont Ronald J.A. Wanders Sacha Ferdinandusse A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl-CoA hydratase deficiency Journal of Lipid Research peroxisomal fatty acid oxidation disorders very long chain fatty acids trihydroxycholestanoic acid |
author_facet |
Jolein Gloerich Simone Denis Elisabeth G. van Grunsven Georges Dacremont Ronald J.A. Wanders Sacha Ferdinandusse |
author_sort |
Jolein Gloerich |
title |
A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl-CoA hydratase deficiency |
title_short |
A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl-CoA hydratase deficiency |
title_full |
A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl-CoA hydratase deficiency |
title_fullStr |
A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl-CoA hydratase deficiency |
title_full_unstemmed |
A novel HPLC-based method to diagnose peroxisomal D-bifunctional protein enoyl-CoA hydratase deficiency |
title_sort |
novel hplc-based method to diagnose peroxisomal d-bifunctional protein enoyl-coa hydratase deficiency |
publisher |
Elsevier |
series |
Journal of Lipid Research |
issn |
0022-2275 |
publishDate |
2003-03-01 |
description |
D-bifunctional protein (D-BP) plays an indispensable role in peroxisomal β-oxidation, and its inherited deficiency in humans is associated with severe clinical abnormalities. Three different subtypes of D-BP deficiency can be distinguished: 1) a complete deficiency of D-BP (type I), 2) an isolated D-BP enoyl-CoA hydratase deficiency (type II), and 3) an isolated D-BP 3-hydroxyacyl-CoA dehydrogenase deficiency (type III). In this study, we developed a method to measure D-BP dehydrogenase activity independent of D-BP hydratase (D-BP HY) activity to distinguish between D-BP deficiency type I and type II, which until now was only possible by mutation analysis. For this assay, the hydratase domain of D-BP was expressed in the yeast Saccharomyces cerevisiae. After a coincubation of yeast homogenate expressing D-BP HY with fibroblast homogenate of patients using the enoyl-CoA ester of the bile acid intermediate trihydroxycholestanoic acid as substrate, D-BP dehydrogenase activity was measured. Fibroblasts of patients with a D-BP deficiency type II displayed D-BP dehydrogenase activity, whereas type I and type III patients did not.This newly developed assay to measure D-BP dehydrogenase activity in fibroblast homogenates provides a quick and reliable method to assign patients with deficient D-BP HY activity to the D-BP deficiency subgroups type I or type II. |
topic |
peroxisomal fatty acid oxidation disorders very long chain fatty acids trihydroxycholestanoic acid |
url |
http://www.sciencedirect.com/science/article/pii/S0022227520312050 |
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