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Understanding pyrroline-5-carboxylate synthetase deficiency: clinical, molecular, functional, and expression studies, structure-based analysis, and novel therapy with arginine

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Journal of Inherited Metabolic Disease

Abstract

Δ1-Pyrroline-5-carboxylate synthetase (P5CS) catalyzes the first two steps of ornithine/proline biosynthesis. P5CS deficiency has been reported in three families, with patients presenting with cutis/joint laxity, cataracts, and neurodevelopmental delay. Only one family exhibited metabolic changes consistent with P5CS deficiency (low proline/ornithine/citrulline/arginine; fasting hyperammonemia). Here we report a new P5CS-deficient patient presenting the complete clinical/metabolic phenotype and carrying p.G93R and p.T299I substitutions in the γ-glutamyl kinase (γGK) component of P5CS. The effects of these substitutions are (1) tested in mutagenesis/functional studies with E.coli γGK, (2) rationalized by structural modelling, and (3) reflected in decreased P5CS protein in patient fibroblasts (shown by immunofluorescence). Using optical/electron microscopy on skin biopsy, we show collagen/elastin fiber alterations that may contribute to connective tissue laxity and are compatible with our angio-MRI finding of kinky brain vessels in the patient. MR spectroscopy revealed decreased brain creatine, which normalized after sustained arginine supplementation, with improvement of neurodevelopmental and metabolic parameters, suggesting a pathogenic role of brain creatine decrease and the value of arginine therapy. Morphological and functional studies of fibroblast mitochondria show that P5CS deficiency is not associated with the mitochondrial alterations observed in Δ1-pyrroline-5-carboxylate reductase deficiency (another proline biosynthesis defect presenting cutis laxa and neurological alterations).

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Acknowledgments

We thank the Association “la Vita è un Dono” for supporting the fellowship of Diego Martinelli, Isabel Pérez-Arellano for advice, and Enrique Pérez-Payá for help with CD analysis. This work was supported by grants BFU2008-05021 of the Spanish Ministry for Science (MEC and MICINN) and Prometeo/2009/051 of the Valencian Government.

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Authors and Affiliations

  1. Division of Metabolism, Bambino Gesù Children’s Hospital, Piazza Sant’Onofrio, 4, 00165, Rome, Italy

    Diego Martinelli, Bianca M. Goffredo, Elsa Bevivino, Sara Boenzi & Carlo Dionisi-Vici

  2. Division of Metabolism and Children’s Research Center (CRC), University Children’s Hospital Zurich, Zurich, Switzerland

    Johannes Häberle, Cecilia Giunta & Matthias R. Baumgartner

  3. Instituto de Biomedicina de Valencia (IBV-CSIC), Valencia, Spain

    Vicente Rubio, Nadine Gougeard & Clara Marco-Marín

  4. Centro de Investigación Biomédica en Red sobre Enfermedades Raras (CIBERER-ISCIII), Valencia, Spain

    Vicente Rubio, Nadine Gougeard & Clara Marco-Marín

  5. Electron Microscopic Laboratory, Dermatological Department and EM Core Facility, University Heidelberg, Heidelberg, Germany

    Ingrid Hausser

  6. Division of Neuromuscular and Neurodegenerative Diseases, Bambino Gesù Children’s Hospital, Rome, Italy

    Rosalba Carrozzo & Maria Chiara Meschini

  7. Divison of Radiology, Bambino Gesù Children’s Hospital, Rome, Italy

    Giovanna Stefania Colafati

  8. Experimental Medicine and Pathology, CSS-Mendel, Rome, Italy

    Francesco Brancati

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  1. Diego Martinelli
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  2. Johannes Häberle
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  3. Vicente Rubio
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  6. Rosalba Carrozzo
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  10. Maria Chiara Meschini
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  14. Francesco Brancati
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  15. Matthias R. Baumgartner
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  16. Carlo Dionisi-Vici
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Corresponding authors

Correspondence to Diego Martinelli or Carlo Dionisi-Vici.

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Communicated by: Jean-Marie Saudubray

Competing interest: None declared

J. Häberle and V. Rubio contributed equally to this work.

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Martinelli, D., Häberle, J., Rubio, V. et al. Understanding pyrroline-5-carboxylate synthetase deficiency: clinical, molecular, functional, and expression studies, structure-based analysis, and novel therapy with arginine. J Inherit Metab Dis 35, 761–776 (2012). https://doi.org/10.1007/s10545-011-9411-8

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