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Detection of 6-demethoxyubiquinone in CoQ10 deficiency disorders: Insights into enzyme interactions and identification of potential therapeutics.
Mol Genet Metab. 2017 07; 121(3):216-223.MG

Abstract

Coenzyme Q10 (CoQ10) is an essential cofactor of the mitochondrial oxidative phosphorylation (OXPHOS) system and its deficiency has important implications for several inherited metabolic disorders of childhood. The biosynthesis of CoQ10 is a complicated process, which involves at least 12 different enzymes. One of the metabolic intermediates that are formed during CoQ10 biosynthesis is the molecule 6-demethoxyubiquinone (6-DMQ). This CoQ precursor is processed at the level of COQ7 and COQ9. We selected this metabolite as a marker substance for metabolic analysis of cell lines with inherited genetic defects (COQ2, COQ4, COQ7 and COQ9) or siRNA knockdown in CoQ biosynthesis enzymes using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). In COQ4, COQ7 and COQ9 deficient cell lines, we detected significantly elevated levels of 6-DMQ. This suggests a functional interplay of these proteins. However, additional siRNA studies demonstrated that elevated 6-DMQ levels are not an exclusive marker of the COQ7/COQ9 enzymatic step of CoQ10 biosynthesis but constitute a more general phenomenon that occurs in disorders impairing the function or stability of the CoQ-synthome. To further investigate the interdependence of CoQ10 biosynthesis enzyme expression, we performed immunoblotting in various cell lines with CoQ10 deficiency, indicating that COQ4, COQ7 and COQ9 protein expression levels are highly regulated depending on the underlying defect. Supplementation of cell lines with synthetic CoQ precursor compounds demonstrated beneficial effects of 2,4-dihydroxybenzoic acid in COQ7 and COQ9 deficiency. Moreover, vanillic acid selectively stimulated CoQ10 biosynthesis and improved cell viability in COQ9 deficiency. However, compounds tested in this study failed to rescue COQ4 deficiency.

Authors+Show Affiliations

Department of General Pediatrics, Neonatology and Pediatric Cardiology, University Children's Hospital, Heinrich-Heine-University Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany.Department of General Pediatrics, Neonatology and Pediatric Cardiology, University Children's Hospital, Heinrich-Heine-University Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany.Institute of Biochemistry, Heinrich-Heine-University, Universitätsstr.1, 40225 Düsseldorf, Germany.Department of General Pediatrics, Neonatology and Pediatric Cardiology, University Children's Hospital, Heinrich-Heine-University Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany.Centre for Inherited Metabolic Diseases, Karolinska University Hospital, Stockholm, Sweden; Max Planck Institute Biology of Ageing - Karolinska Institutet Laboratory, Division of Metabolic Diseases, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden.Institute of Human Genetics, Technische Universität München, Trogerstr. 32, 81675 Munich, Germany; Institute of Human Genetics, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.Clinic for Pediatrics, Division of Inherited Metabolic Disorders, Medical University of Innsbruck, 6020 Innsbruck, Austria.Centre for Inherited Metabolic Diseases, Karolinska University Hospital, Stockholm, Sweden; Max Planck Institute Biology of Ageing - Karolinska Institutet Laboratory, Division of Metabolic Diseases, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden.Centre for Inherited Metabolic Diseases, Karolinska University Hospital, Stockholm, Sweden; Max Planck Institute Biology of Ageing - Karolinska Institutet Laboratory, Division of Metabolic Diseases, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; Department of Molecular Medicine and Surgery, Science for Life Laboratory, Karolinska Institute, Stockholm, Sweden.Departmento de Fisiología, Facultad de Medicina and Instituto de Biotecnología, Centro de Investigación Biomédica, Universidad de Granada, Spain.Department of General Pediatrics, Neonatology and Pediatric Cardiology, University Children's Hospital, Heinrich-Heine-University Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany.Department of General Pediatrics, Neonatology and Pediatric Cardiology, University Children's Hospital, Heinrich-Heine-University Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany. Electronic address: Felix.distelmaier@med.uni-duesseldorf.de.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

28552678

Citation

Herebian, Diran, et al. "Detection of 6-demethoxyubiquinone in CoQ10 Deficiency Disorders: Insights Into Enzyme Interactions and Identification of Potential Therapeutics." Molecular Genetics and Metabolism, vol. 121, no. 3, 2017, pp. 216-223.
Herebian D, Seibt A, Smits SHJ, et al. Detection of 6-demethoxyubiquinone in CoQ10 deficiency disorders: Insights into enzyme interactions and identification of potential therapeutics. Mol Genet Metab. 2017;121(3):216-223.
Herebian, D., Seibt, A., Smits, S. H. J., Bünning, G., Freyer, C., Prokisch, H., Karall, D., Wredenberg, A., Wedell, A., López, L. C., Mayatepek, E., & Distelmaier, F. (2017). Detection of 6-demethoxyubiquinone in CoQ10 deficiency disorders: Insights into enzyme interactions and identification of potential therapeutics. Molecular Genetics and Metabolism, 121(3), 216-223. https://doi.org/10.1016/j.ymgme.2017.05.012
Herebian D, et al. Detection of 6-demethoxyubiquinone in CoQ10 Deficiency Disorders: Insights Into Enzyme Interactions and Identification of Potential Therapeutics. Mol Genet Metab. 2017;121(3):216-223. PubMed PMID: 28552678.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of 6-demethoxyubiquinone in CoQ10 deficiency disorders: Insights into enzyme interactions and identification of potential therapeutics. AU - Herebian,Diran, AU - Seibt,Annette, AU - Smits,Sander H J, AU - Bünning,Gisela, AU - Freyer,Christoph, AU - Prokisch,Holger, AU - Karall,Daniela, AU - Wredenberg,Anna, AU - Wedell,Anna, AU - López,Luis C, AU - Mayatepek,Ertan, AU - Distelmaier,Felix, Y1 - 2017/05/20/ PY - 2017/02/13/received PY - 2017/05/18/revised PY - 2017/05/19/accepted PY - 2017/5/30/pubmed PY - 2018/3/20/medline PY - 2017/5/30/entrez KW - 2,4-Dihydroxybenzoic acid KW - 6-DMQ KW - Coenzyme Q(10) KW - Mitochondria KW - Vanillic acid SP - 216 EP - 223 JF - Molecular genetics and metabolism JO - Mol Genet Metab VL - 121 IS - 3 N2 - Coenzyme Q10 (CoQ10) is an essential cofactor of the mitochondrial oxidative phosphorylation (OXPHOS) system and its deficiency has important implications for several inherited metabolic disorders of childhood. The biosynthesis of CoQ10 is a complicated process, which involves at least 12 different enzymes. One of the metabolic intermediates that are formed during CoQ10 biosynthesis is the molecule 6-demethoxyubiquinone (6-DMQ). This CoQ precursor is processed at the level of COQ7 and COQ9. We selected this metabolite as a marker substance for metabolic analysis of cell lines with inherited genetic defects (COQ2, COQ4, COQ7 and COQ9) or siRNA knockdown in CoQ biosynthesis enzymes using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). In COQ4, COQ7 and COQ9 deficient cell lines, we detected significantly elevated levels of 6-DMQ. This suggests a functional interplay of these proteins. However, additional siRNA studies demonstrated that elevated 6-DMQ levels are not an exclusive marker of the COQ7/COQ9 enzymatic step of CoQ10 biosynthesis but constitute a more general phenomenon that occurs in disorders impairing the function or stability of the CoQ-synthome. To further investigate the interdependence of CoQ10 biosynthesis enzyme expression, we performed immunoblotting in various cell lines with CoQ10 deficiency, indicating that COQ4, COQ7 and COQ9 protein expression levels are highly regulated depending on the underlying defect. Supplementation of cell lines with synthetic CoQ precursor compounds demonstrated beneficial effects of 2,4-dihydroxybenzoic acid in COQ7 and COQ9 deficiency. Moreover, vanillic acid selectively stimulated CoQ10 biosynthesis and improved cell viability in COQ9 deficiency. However, compounds tested in this study failed to rescue COQ4 deficiency. SN - 1096-7206 UR - https://www.unboundmedicine.com/medline/citation/28552678/Detection_of_6_demethoxyubiquinone_in_CoQ10_deficiency_disorders:_Insights_into_enzyme_interactions_and_identification_of_potential_therapeutics_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1096-7192(17)30100-2 DB - PRIME DP - Unbound Medicine ER -