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Inhibition of 3-methylcrotonyl-CoA carboxylase explains the increased excretion of 3-hydroxyisovaleric acid in valproate-treated patients

  • Original Article
  • Published:
Journal of Inherited Metabolic Disease

Abstract

Background

Valproic acid (VPA) is a widely used anticonvulsant drug which affects mitochondrial metabolism including the catabolism of fatty acids and branched-chain amino acids.

Aims

To elucidate the effect of valproate on the leucine pathway through a targeted metabolomics approach and the evaluation of the effects of valproate on the activity of biotinidase and 3-methylcrotonyl-CoA carboxylase (3MCC).

Methods

Urine organic acid analysis was performed in patients under VPA therapy and healthy controls using gas-chromatography/mass spectrometry (GC-MS). Biotinidase activity was determined in plasma samples of both groups using an optimized spectrophotometric assay. After immunoprecipitation of short-chain enoyl-CoA hydratase (crotonase, ECHS1), 3MCC activity was measured in human liver homogenate using high-performance liquid chromatography (HPLC), in the absence and presence of valproyl-CoA.

Results

The levels of 3-hydroxyisovaleric acid (3OH-IVA), one secondary metabolite of the leucine pathway, were significantly elevated in human urine after VPA treatment. Biotinidase activity in plasma samples ranged from very low to normal levels in treated patients as compared with controls. Enzyme activity measurements revealed inhibition of 3-methylcrotonyl-CoA carboxylase by valproyl-CoA (IC50 = 1.36 mM). Furthermore, we show that after complete immunoprecipitation of crotonase in a human liver homogenate, 3-hydroxyisovaleryl-CoA is not formed.

Discussion

Our results suggest the interference of VPA with the activity of 3MCC through a potential cumulative effect: direct inhibition of the enzyme activity by the drug metabolite valproyl-CoA and the inhibition of biotinidase by valproate and/or its metabolites. These interactions may be associated with the skin rash and hair loss which are side effects often reported in VPA-treated patients.

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Abbreviations

VPA:

2-n-propylpentanoic acid or valproic acid

CoA:

Coenzyme A

VP-CoA:

Valproyl-CoA

3OH-IVA:

3-hydroxyisovaleric acid

3MCC:

3-Methylcrotonyl-CoA carboxylase (EC 6.4.1.4)

ECHS1:

Short-chain enoyl-CoA hydratase or crotonase (EC 4.2.1.17)

BTD:

Biotinidase (EC 3.5.1.12)

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Acknowledgements

This work was financially supported by Fundação para a Ciência e a Tecnologia (FCT), Lisboa, Portugal (SFRH/BD/25913/2005).

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Authors and Affiliations

  1. Research Institute for Medicines and Pharmaceutical Sciences, iMed.UL, Faculdade de Farmácia da Universidade de Lisboa, Lisboa, Portugal

    Paula B. M. Luís, Isabel Tavares de Almeida & Margarida F. B. Silva

  2. Department of Biochemistry and Human Biology, Faculdade de Farmácia da Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003, Lisboa, Portugal

    Isabel Tavares de Almeida & Margarida F. B. Silva

  3. Department of Clinical Chemistry and Pediatrics, Academic Medical Center, Amsterdam, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands

    Jos P. Ruiter, Lodewijk IJlst, Marinus Duran & Ronald J. Wanders

  4. Centro de Desenvolvimento Luís Borges, Hospital Pediátrico de Coimbra, Coimbra, Portugal

    Luísa Diogo & Paula Garcia

Authors
  1. Paula B. M. Luís
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  2. Jos P. Ruiter
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  3. Lodewijk IJlst
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  4. Luísa Diogo
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  6. Isabel Tavares de Almeida
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  7. Marinus Duran
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  8. Ronald J. Wanders
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  9. Margarida F. B. Silva
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Corresponding author

Correspondence to Margarida F. B. Silva.

Additional information

Communicated by: K. Michael Gibson

Ronald J. Wanders and Margarida F. B. Silva are equal last authors

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Luís, P.B.M., Ruiter, J.P., IJlst, L. et al. Inhibition of 3-methylcrotonyl-CoA carboxylase explains the increased excretion of 3-hydroxyisovaleric acid in valproate-treated patients. J Inherit Metab Dis 35, 443–449 (2012). https://doi.org/10.1007/s10545-011-9423-4

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  • DOI: https://doi.org/10.1007/s10545-011-9423-4

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