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JIMD Reports, Volume 37 pp 13–17Cite as

Guanidinoacetate Methyltransferase Activity in Lymphocytes, for a Fast Diagnosis

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Part of the book series: JIMD Reports ((JIMD,volume 37))

Abstract

Introduction. Guanidinoacetate methyltransferase (GAMT) deficiency is an inborn error of metabolism (IEM), clinically characterized by intellectual disability, developmental delay, seizures, and movement disorders. Biochemical diagnosis of GAMT deficiency is based on the measurement of creatine and guanidinoacetate in urine, plasma, or CSF and is confirmed genetically by DNA analysis or by enzyme assay in lymphoblasts or fibroblasts. To obtain enough cells, these cells need to be cultured for at least 1 month. A less time-consuming diagnostic functional test is needed, since GAMT deficiency is a candidate for newborn screening (NBS) programs, to be able to confirm or rule out this IEM after an initial positive result in the NBS.

Methods. Stable-isotope-labeled 13C2-guanidinoacetate and 2H3-S-adenosylmethionine (SAM) were used, which are converted by GAMT present in lymphocyte extracts into 2H3-13C2-creatine. The formed 2H3-13C2-creatine was butylated and subsequently measured by liquid chromatography tandem mass-spectrometry (LC-MS/MS).

Results. We measured GAMT enzyme activity in lymphocyte extracts of 24 controls, 3 GAMT deficient patients and of 2 parents proven to be carrier. Because GAMT activity decreases when isolation time after venipuncture increases, reference values were obtained for 2 control groups: isolation on the day of venipuncture (27–130 pmol/h/mg) and 1 day afterwards (15–146 pmol/h/mg). Deficient patients had no detectable GAMT activity. The two carriers had GAMT activity within the normal range.

Conclusion. We designed a fast, less invasive, and valid method to measure GAMT activity in lymphocytes using LC-MS/MS analysis without the need of time-consuming and laborious cell culture.

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Abbreviations

AGAT:

Arginine:glycine amidinotransferase

EDTA:

Ethylenediaminetetraacetic acid

GAMT:

Guanidinoacetate methyltransferase

HBSS:

Hanks balanced salt solution

LC-MS/MS:

Liquid-chromatography tandemmass-spectrometry

MRS:

Magnetic resonance spectroscopy

NBS:

Newborn screening

SAM:

S-adenosylmethionine

tris:

Tris(hydroxymethyl)aminomethane

WES:

Whole exome sequencing

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Authors and Affiliations

  1. Metabolic Laboratory, Department of Clinical Chemistry, Amsterdam Neuroscience, VU Medical Center, Amsterdam, The Netherlands

    Lisette M. Berends, Eduard A. Struys, Birthe Roos, Ulbe Holwerda, Erwin E. W. Jansen, Gajja S. Salomons & Mirjam M. C. Wamelink

Authors
  1. Lisette M. Berends
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  2. Eduard A. Struys
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  3. Birthe Roos
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  4. Ulbe Holwerda
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  5. Erwin E. W. Jansen
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  6. Gajja S. Salomons
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  7. Mirjam M. C. Wamelink
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Corresponding author

Correspondence to Mirjam M. C. Wamelink .

Editor information

Editors and Affiliations

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

    Eva Morava

  2. Division of Metabolism & Children's Research Centre, University Children's Hospital Zürich, Zürich, 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: Brian Fowler, PhD

Appendices

Synopsis

A new, fast, less invasive and valid method to measure GAMT activity in lymphocytes using LC-MS/MS analysis of a 2H3-13C2-creatine butyl derivative was designed.

Compliance with Ethics Guidelines

Lisette M. Berends, Eduard A. Struys, Birthe Roos, Ulbe Holwerda, Erwin E. W. Jansen, Gajja S. Salomons, and Mirjam M. C. Wamelink declare that they have no conflict of interest.

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000 (5). Informed consent was obtained from all patients for being included in the study.

Author Contribution

Lisette M. Berends: conception and design; analysis and interpretation of data; drafting the article; revising it critically.

Eduard. A. Struys: analysis and interpretation of data; revising it critically.

Birthe Roos: conception and design; analysis and interpretation of data; revising it critically.

Ulbe Holwerda: analysis and interpretation of data; revising it critically.

Erwin E. W. Jansen: analysis and interpretation of data; revising it critically.

Gajja S. Salomons: conception and design; revising it critically.

Mirjam M. C. Wamelink: conception and design; analysis and interpretation of data; revising it critically, guarantor.

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© 2017 SSIEM and Springer-Verlag Berlin Heidelberg

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Berends, L.M. et al. (2017). Guanidinoacetate Methyltransferase Activity in Lymphocytes, for a Fast Diagnosis. In: Morava, E., Baumgartner, M., Patterson, M., Rahman, S., Zschocke, J., Peters, V. (eds) JIMD Reports, Volume 37. JIMD Reports, vol 37. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8904_2017_4

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  • DOI: https://doi.org/10.1007/8904_2017_4

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  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-662-56358-8

  • Online ISBN: 978-3-662-56359-5

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

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