Chapter title |
Radioligand Binding Assays for Determining Dissociation Constants of Phytohormone Receptors
|
---|---|
Chapter number | 3 |
Book title |
Plant Proteostasis
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3759-2_3 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3757-8, 978-1-4939-3759-2
|
Authors |
Antje Hellmuth, Luz Irina A. Calderón Villalobos |
Editors |
L. Maria Lois, Rune Matthiesen |
Abstract |
In receptor-ligand interactions, dissociation constants provide a key parameter for characterizing binding. Here, we describe filter-based radioligand binding assays at equilibrium, either varying ligand concentrations up to receptor saturation or outcompeting ligand from its receptor with increasing concentrations of ligand analogue. Using the auxin coreceptor system, we illustrate how to use a saturation binding assay to determine the apparent dissociation constant (K D (') ) for the formation of a ternary TIR1-auxin-AUX/IAA complex. Also, we show how to determine the inhibitory constant (K i) for auxin binding by the coreceptor complex via a competition binding assay. These assays can be applied broadly to characterize a one-site binding reaction of a hormone to its receptor. |
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Demographic breakdown
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Members of the public | 1 | 100% |
Mendeley readers
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Unknown | 11 | 100% |
Demographic breakdown
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Student > Bachelor | 3 | 27% |
Student > Master | 2 | 18% |
Other | 1 | 9% |
Researcher | 1 | 9% |
Unknown | 4 | 36% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 2 | 18% |
Agricultural and Biological Sciences | 1 | 9% |
Chemistry | 1 | 9% |
Unknown | 4 | 36% |