Skip to main content
SpringerLink
Log in

Development of a Tandem Mass Spectrometry Method for Rapid Measurement of Medium- and Very-Long-Chain Acyl-CoA Dehydrogenase Activity in Fibroblasts

  • Research Report
  • Chapter
  • First Online:
JIMD Reports, Volume 35

Part of the book series: JIMD Reports ((JIMD,volume 35))

Abstract

Mitochondrial fatty acid oxidation is a vital biochemical process for energy metabolism. Among the known fatty-acid metabolism disorders, very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency and medium-chain acyl-CoA dehydrogenase (MCAD) deficiency count among the most frequent. Both are potentially very serious diseases as they carry a risk of severe neurological post-crisis sequelae, and even sudden death. Diagnosis relies on plasma acylcarnitine profile analysis and urine organic acid analysis, followed by genetic testing to confirm diagnosis. However, in some cases, it is crucial to run a specific diagnostic assay for enzyme activity, which is generally performed in leukocytes or fibroblasts. The aim of this study was to address this need, first by developing a MCAD and VLCAD enzyme activity-specific diagnostic assay in fibroblasts (by measuring the reaction products, i.e. enoyl-CoA) via a rapid LC–MS/MS-based technique, and then by testing MCAD-deficient patients (n = 6), VLCAD-deficient patients (n = 10), and control patients (n = 12). MCAD activity was significantly different in the MCAD-deficiency (MCADD) group (mean = 0.07 nmol C8:1 formed/min/mg protein) compared to the control group (mean = 0.36 nmol C8:1 formed/min/mg protein). All MCADD patients showed less than 35% residual MCAD activity. VLCAD activity was significantly decreased in the VLCADD group (mean = 0.06 nmol C16:1 formed/min/mg protein) compared to the control group (mean = 0.86 nmol C16:1 formed/min/mg protein, respectively). All VLCADD patients showed less than 35% residual VLCAD activity. This technique allowed also to confirm that a novel ACADVL gene mutation (c.1400T>C) is responsible for a defective VLCAD activity (residual activity at 10%).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Abbreviations

ETF:

Electron Transfer Flavoprotein

FA:

Fatty acids

FAD:

Flavin adenine dinucleotide

HPLC:

High-performance liquid chromatography

MCAD:

Medium-chain acyl-CoA dehydrogenase

MCADD:

MCAD deficiency

MES:

2-(N-morpholino)ethanesulfonic acid

OMIM:

Online Mendelian Inheritance in Man

VLCAD:

Very-long-chain acyl-CoA dehydrogenase

VLCADD:

VLCAD deficiency

References

  • Arnold GL, Van Hove J, Freedenberg D et al (2009) A Delphi clinical practice protocol for the management of very long chain acyl-CoA dehydrogenase deficiency. Mol Genet Metab 96(3):85–90

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bertrand C, Dumoulin R, Divry P, Mathieu M, Vianey-Saban C (1992) Purification of electron transfer flavoprotein from pig liver mitochondria and its application to the diagnosis of deficiencies of acyl-CoA dehydrogenases in human fibroblasts. Clin Chim Acta 210(1-2):75–91

    Article  CAS  PubMed  Google Scholar 

  • Feillet F, Ogier H, Cheillan D et al (2012) Medium-chain acyl-CoA-dehydrogenase (MCAD) deficiency: french consensus for neonatal screening, diagnosis, and management. Arch Pediatr 19(2):184–193

    Article  CAS  PubMed  Google Scholar 

  • Hashimoto T (1992) Peroxisomal and mitochondrial enzymes. Prog Clin Biol Res 375:19–32

    CAS  PubMed  Google Scholar 

  • Houten SM, Wanders RJA (2010) A general introduction to the biochemistry of mitochondrial fatty acid β-oxidation. J Inherit Metab Dis 33:469–477

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McKinney JT, Longo N, Hahn SH et al (2004) Rapid, comprehensive screening of the human medium chain acyl-CoA dehydrogenase gene. Mol Genet Metab 82(2):112–120

    Article  CAS  PubMed  Google Scholar 

  • Souri M, Aoyama T, Orii K, Yamaguchi S, Hashimoto T (1996) Mutation analysis of very-long-chain acyl-coenzyme A dehydrogenase (VLCAD) deficiency: identification and characterization of mutant VLCAD cDNAs from four patients. Am J Hum Genet 58(1):97–106

    CAS  PubMed  PubMed Central  Google Scholar 

  • Tajima G, Sakura N, Shirao K et al (2008) Development of a new enzymatic diagnosis method for very-long-chain Acyl-CoA dehydrogenase deficiency by detecting 2-hexadecenoyl-CoA production and its application in tandem mass spectrometry-based selective screening and newborn screening in Japan. Pediatr Res 64(6):667–672

    Article  CAS  PubMed  Google Scholar 

  • ter Veld F, Mueller M, Kramer S et al (2009) A novel tandem mass spectrometry method for rapid confirmation of medium- and very long-chain acyl-CoA dehydrogenase deficiency in newborns. PLoS One 4:e6449

    Article  PubMed  PubMed Central  Google Scholar 

  • Vianey-Saban C, Divry P, Brivet M et al (1998) Mitochondrial very-long-chain acyl-coenzyme A dehydrogenase deficiency: clinical characteristics and diagnostic considerations in 30 patients. Clin Chim Acta 269(1):43–62

    Article  CAS  PubMed  Google Scholar 

  • Waddell L, Wiley V, Carpenter K et al (2006) Medium-chain acyl-CoA dehydrogenase deficiency: genotype-biochemical phenotype correlations. Mol Genet Metab 87(1):32–39

    Article  CAS  PubMed  Google Scholar 

  • Wanders RJ, Vreken P, den Boer ME, Wijburg FA, van Gennip AH, IJlst L (1999) Disorders of mitochondrial fatty acyl-CoA beta-oxidation. J Inherit Metab Dis 22(4):442–487

    Article  CAS  PubMed  Google Scholar 

  • Wanders RJ, Ruiter JP, IJLst L, Waterham HR, Houten SM (2010) The enzymology of mitochondrial fatty acid beta-oxidation and its application to follow-up analysis of positive neonatal screening results. J Inherit Metab Dis 33(5):479–494

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

The authors thank A.T.T. (Auvergne Traduction Technique®) for proofreading the manuscript.

Author information

Authors and Affiliations

  1. Service de Biochimie Médicale, Centre de Biologie, CHU Gabriel-Montpied, rue Montalembert, 63000, Clermont-Ferrand, France

    Damien Bouvier

  2. Retinoids, Reproduction Developmental Diseases, School of Medicine, Clermont Université, Université d’Auvergne, EA7281, 63000, Clermont-Ferrand, France

    Damien Bouvier

  3. Service Maladies Héréditaires du Métabolisme et Dépistage Néonatal, Centre de Biologie et de Pathologie Est, CHU Lyon, INSERM U1060 CarMeN, 69500, Bron, France

    Christine Vianey-Saban & Séverine Ruet

  4. Service Maladies Héréditaires du Métabolisme et Dépistage Néonatal, Centre de Biologie et de Pathologie Est, CHU Lyon, UMR 5305 CNRS/UCBL, 69500, Bron, France

    Cécile Acquaviva

Authors
  1. Damien Bouvier
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christine Vianey-Saban
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Séverine Ruet
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cécile Acquaviva
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cécile Acquaviva .

Editor information

Editors and Affiliations

  1. Tulane University Medical School , New Orleans, Louisiana, USA

    Eva Morava

  2. Division of Metabolism and Children’s Research Centre, University Children’s Hospital Zurich, Zurich, Switzerland

    Matthias Baumgartner

  3. Division of Child and Adolescent Neurology, Mayo Clinic, Rochester, Minnesota, USA

    Marc Patterson

  4. Clinical and Molecular Genetics Unit, UCL Institute of Child Health, London, United Kingdom

    Shamima Rahman

  5. Division of Human Genetics, Medical University Innsbruck, Innsbruck, Austria

    Johannes Zschocke

  6. Center for Child and Adolescent Medicine, Heidelberg University Hospital, Heidelberg, Germany

    Verena Peters

Additional information

Communicated by: Cornelis Jakobs

Appendices

Take-Home Message

The reader will learn about the usefulness of MCAD and VLCAD activity measurement and the performance of the method developed in fibroblasts.

Conflict of Interest

Damien Bouvier, Christine Vianey-Saban, Séverine Ruet, and Cécile Acquaviva declare that they have no conflict of interest.

Compliance with ethics guidelines: Informed consent was obtained from all patients for being included in the study.

Authors Contribution

Damien Bouvier and Severine Ruet have performed technical part of the study.

Damien Bouvier, Christine Vianey-Saban, Severine Ruet, and Cécile Acquaviva have performed analysis and interpretation of the data.

Damien Bouvier drafted the article.

Christine Vianey-Saban and Cécile Acquaviva revised the article.

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Society for the Study of Inborn Errors of Metabolism (SSIEM)

About this chapter

Cite this chapter

Bouvier, D., Vianey-Saban, C., Ruet, S., Acquaviva, C. (2016). Development of a Tandem Mass Spectrometry Method for Rapid Measurement of Medium- and Very-Long-Chain Acyl-CoA Dehydrogenase Activity in Fibroblasts. In: Morava, E., Baumgartner, M., Patterson, M., Rahman, S., Zschocke, J., Peters, V. (eds) JIMD Reports, Volume 35. JIMD Reports, vol 35. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8904_2016_22

Download citation

  • DOI: https://doi.org/10.1007/8904_2016_22

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-662-55832-4

  • Online ISBN: 978-3-662-55833-1

  • eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)

Publish with us

Policies and ethics

Search

Navigation