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Quantitation of Glutamine Synthetase 1 Activity in Drosophila melanogaster

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Metabolic Reprogramming

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2675))

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Abstract

Protocols to assay the activity of glutamine synthetase (GS) are presented as they have been used in our laboratory to correlate the expression levels of the gene encoding Drosophila GS1 gene, the GS1 protein levels, and its activity in extracts of larvae and heads from Drosophila melanogaster. The assays are based on the glutamine synthetase-catalyzed formation of γ-glutamylhydroxylamine in the presence of ATP, L-glutamate, and hydroxylamine, in which hydroxylamine substitutes for ammonia in the reaction. Formation of γ-glutamylhydroxylamine is monitored spectrophotometrically in discontinuous assays upon complex formation with FeCl3. Fixed-time assays and those based on monitoring the time-course of product formation at different reaction times are described. The protocols can be adapted to quantify glutamine synthetase activity on biological materials other than Drosophila.

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Acknowledgments

This research was funded by the Cariplo Foundation, grant n. 20140703, and the European Huntington’s Disease Network seed funds n. 689 to P.B.

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Authors and Affiliations

  1. Dipartimento di Bioscienze, Università degli Studi di Milano, Milan, Italy

    Teresa Vitali & Maria Antonietta Vanoni

  2. Department of Anatomy and Cell Biology, George Washington University School of Medicine and Health Sciences, Washington, DC, USA

    Teresa Vitali

  3. Department of Cellular, Computational and Integrative Biology (CIBIO), University of Trento, Trento, Italy

    Paola Bellosta

  4. Department of Medicine, New York University-Langone Medical Center, New York, NY, USA

    Paola Bellosta

Authors
  1. Teresa Vitali
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  2. Maria Antonietta Vanoni
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  3. Paola Bellosta
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Corresponding authors

Correspondence to Maria Antonietta Vanoni or Paola Bellosta .

Editor information

Editors and Affiliations

  1. Leeds Institute of Medical Research, St. James’s University Hospital, University of Leeds, Leeds, UK

    Salvatore Papa

  2. Center for Genome Engineering and Maintenance, Department of Life Sciences, College of Health, Medicine and Life Sciences, Brunel University London, London, UK

    Concetta Bubici

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Vitali, T., Vanoni, M.A., Bellosta, P. (2023). Quantitation of Glutamine Synthetase 1 Activity in Drosophila melanogaster. In: Papa, S., Bubici, C. (eds) Metabolic Reprogramming. Methods in Molecular Biology, vol 2675. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3247-5_18

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  • DOI: https://doi.org/10.1007/978-1-0716-3247-5_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3246-8

  • Online ISBN: 978-1-0716-3247-5

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