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Effect of Cysteamine on Mutant ASL Proteins with Cysteine for Arginine Substitutions

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Abstract

Introduction

Cysteamine is used to treat cystinosis via the modification of cysteine residues substituting arginine in mutant proteins.

Objectives

We investigated the effect of cysteamine on mutant argininosuccinate lyase (ASL), the second most common defect in the urea cycle.

Methods

In an established mammalian expression system, 293T cell lysates were produced after transfection with all known cysteine for arginine mutations in the ASL gene (p.Arg94Cys, p.Arg95Cys, p.Arg168Cys, p.Arg379Cys, and p.Arg385Cys), allowing testing of the effect of cysteamine over 48 h in the culture medium as well as for 1 h immediately prior to the enzyme assay.

Results

Cysteamine at low concentrations showed no effect on 293T cell viability, ASL protein expression, or ASL activity when applied during cell culture. However, incubation of transfected cells with 0.05 mM cysteamine immediately before the enzyme assay resulted in increased ASL activity of p.Arg94Cys, p.Arg379Cys, and p.Arg385Cys by 64, 20, and 197 %, respectively, and this result was significant (p < 0.01). Cell lysates carrying p.Arg385Cys and treated with cysteamine recover enzyme activity that is similar to the untreated designed mutation p.Arg385Lys, providing circumstantial evidence for the assumed cysteamine-induced change of a cysteine to a lysine analogue.

Conclusion

Since 12 % of all known genotypes in ASL deficiency are affected by a cysteine for arginine mutation, we conclude that the potential of cysteamine or of related substances as remedy for this disease should be investigated further.

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Authors and Affiliations

  1. Division of Metabolism and Children’s Research Centre (CRC), University Children’s Hospital Zürich, Steinwiesstr. 75, 8032, Zurich, Switzerland

    Corinne Inauen, Véronique Rüfenacht, Liyan Hu & Johannes Häberle

  2. Pediatric Endocrinology, University Children’s Hospital and Department of Clinical Research, University of Bern, Bern, Switzerland

    Amit V. Pandey

  3. Affiliated with the Neuroscience Center Zürich (ZNZ), Zurich, Switzerland

    Liyan Hu & Johannes Häberle

  4. Department of General Pediatrics, Adolescent Medicine and Neonatology, University Medical Centre Freiburg, Freiburg, Germany

    Henk Blom

  5. University Institute of Clinical Chemistry, University of Bern, Bern, Switzerland

    Jean-Marc Nuoffer

  6. University Children’s Hospital, University of Bern, Bern, Switzerland

    Jean-Marc Nuoffer

  7. Affiliated with the Zürich Center for Integrative Human Physiology (ZIHP), Zurich, Switzerland

    Johannes Häberle

Authors
  1. Corinne Inauen
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  2. Véronique Rüfenacht
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  3. Amit V. Pandey
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  4. Liyan Hu
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  5. Henk Blom
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  6. Jean-Marc Nuoffer
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  7. Johannes Häberle
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Corresponding author

Correspondence to Johannes Häberle.

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Conflict of interest

The authors CI, VR, AVP, LH, HB, JMN, and JH declare that they have no conflicts of interest.

Funding

This work was supported by the Swiss National Science Foundation (Grants 310030_153196/1 to JH and 310031_134926 to AVP) and a grant from Schweizerische Mobiliar Genossenschaft Jubiläumsstiftung to AVP.

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Inauen, C., Rüfenacht, V., Pandey, A.V. et al. Effect of Cysteamine on Mutant ASL Proteins with Cysteine for Arginine Substitutions. Mol Diagn Ther 20, 125–133 (2016). https://doi.org/10.1007/s40291-015-0182-z

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  • DOI: https://doi.org/10.1007/s40291-015-0182-z

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