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Conventional Receptor Radioligand Binding Techniques Applied to the Study of Monoamine Oxidase

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Monoamine Oxidase

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2558))

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Abstract

Designed to measure binding interactions between small molecules and receptor proteins, radioligand binding approaches may also be applied to interactions between monoamine oxidase (MAO) and its ligands. The technique may be used with tissue homogenates or with mitochondrial membranes and can provide information about binding site density, ligand affinity, binding rate constants, and binding events at sites that do not impact absorbance characteristics of the flavin cofactor and that may not be amenable to spectrophotometric studies. This overview describes the use of a cell harvester in a common filtration approach to measure binding to MAO of radiolabeled substrates, inhibitors, or allosteric ligands in saturation analyses and to take advantage of the principles of competition to obtain quantitative binding data for unlabeled ligands that may bind with much lower affinity. The quality and reproducibility of data are impacted by factors such as choice of ligand concentrations, pipetting technique, graphing and regression approaches, and scintillation counting parameters, and consideration is given to these and other factors that may influence the results.

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Acknowledgements

I thank Professor David Colquhoun for helpful discussions.

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Authors and Affiliations

  1. Neurochemical Research Unit, Department of Psychiatry, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, Canada

    Andrew Holt

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  1. Andrew Holt
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Correspondence to Andrew Holt .

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Editors and Affiliations

  1. Department of Biology and Biotechnology, University of Pavia, Pavia, Italy

    Claudia Binda

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Holt, A. (2023). Conventional Receptor Radioligand Binding Techniques Applied to the Study of Monoamine Oxidase. In: Binda, C. (eds) Monoamine Oxidase. Methods in Molecular Biology, vol 2558. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2643-6_7

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  • DOI: https://doi.org/10.1007/978-1-0716-2643-6_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2642-9

  • Online ISBN: 978-1-0716-2643-6

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