ADRENERGIC RECEPTORS

Crystallization and structure determination of adrenergic G protein-coupled receptor subtypes

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 Obiettivo del progetto (Objective)

'Crystal structures of beta1 and beta2 adrenergic G protein-coupled receptors (GPCRs) have been determined recently. We propose to extend the structural information about this physiologically and medically important GPCR subfamily by obtaining the structures of the beta3 and the alpha adrenergic receptors. The ample experience in GPCR structural biology of the host laboratory, complemented by its multidisciplinary approach, provides an ideal environment for this challenging task. We will make use of high-throughput facilities and methods to optimize the receptor constructs for expression and protein stability. We will use various crystallization methods and available robotic nano-liter dispensers to determine suitable conditions for crystal growth. We will have access to state-of-the-art microcrystallography beamlines at the Swiss Light Source synchrotron at Paul Scherrer Institut (PSI) to test a large number of obtained crystals and collect the best possible diffraction datasets. Once diffraction data of sufficient quality has been obtained, the structure will be solved by molecular replacement. In collaboration with other PSI scientists and external academic and industrial partners we will use the structural results obtained for the adrenergic receptor subtypes to study the structural basis of ligand efficacy, i.e. how the process of ligand binding is translated into receptor function, and to find applications in therapeutic drug development.'

Introduzione (Teaser)

A European research group is working to obtain the crystal structure of adrenergic receptors. Knowing the three-dimensional structure of the receptor could help in designing drugs to block or activate its function.

Descrizione progetto (Article)

Adrenergic receptors are expressed by various cell types and are central for human physiology. In humans, there are nine subtypes of adrenergic receptors with different functions and pharmacology.

Given that adrenergic receptors are important drug targets for treating various conditions, structural studies will help scientists unveil their distinct pharmacological properties. However, the three-dimensional structure of beta 1 and beta 2 receptors has only recently been elucidated. The EU-funded 'Crystallization and structure determination of adrenergic G protein-coupled receptor subtypes' (ADRENERGIC RECEPTORS) project proposes to extend this data and obtain the crystal structure of other beta and alpha adrenergic receptors.

To obtain stable receptors suitable for crystallisation procedures, researchers are expressing thermostable mutants or fusing receptors with small soluble proteins. These receptor-expressing constructs are introduced into mammalian or insect cells and the modified receptors are purified using affinity chromatography methods.

The ongoing crystallisation experiments are also performed with the addition of antagonist molecules. Subsequent analysis with nuclear magnetic resonance spectroscopy should provide insight into the structural basis of ligand selectivity. Moreover, this enables the study of the dynamic changes in the receptors upon ligand binding.

Given the difficulties associated with the crystallisation process, the ADRENERGIC RECEPTORS work constitutes advancement in the technology. In addition, it opens up avenues for using the 3D structure of medically relevant receptors to design effective drugs, hopefully with fewer side-effects.