* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download G-protein coupled receptor over-expression in
Phosphorylation wikipedia , lookup
Cytokinesis wikipedia , lookup
Theories of general anaesthetic action wikipedia , lookup
Cell membrane wikipedia , lookup
Protein (nutrient) wikipedia , lookup
Magnesium transporter wikipedia , lookup
Endomembrane system wikipedia , lookup
Protein phosphorylation wikipedia , lookup
Homology modeling wikipedia , lookup
Protein domain wikipedia , lookup
Protein moonlighting wikipedia , lookup
Protein structure prediction wikipedia , lookup
Intrinsically disordered proteins wikipedia , lookup
Nuclear magnetic resonance spectroscopy of proteins wikipedia , lookup
Signal transduction wikipedia , lookup
List of types of proteins wikipedia , lookup
Protein–protein interaction wikipedia , lookup
Understanding structure-activity relationships in G-protein coupled receptors (GPCRs) Dr Roslyn Bill and Dr David Poyner http://www.aston.ac.uk/lhs/staff/A-Zindex/billrm.jsp http://www.aston.ac.uk/lhs/staff/A-Zindex/poynerdr.jsp GPCRs are the single largest protein family in the mammalian genome, and the largest class of drug targets. Unfortunately, they are only available in minute quantities in the cell (typically less than 0.1% of the protein complement). It is therefore recognised by the scientific community that the only way to overcome the bottleneck in gaining a complete understanding of GPCRs and thus to develop new drugs is to produce sufficient quantities of this key class of proteins for structural and functional analysis. Currently, the only available high-resolution structure of a GPCR is that of rhodopsin, the chief vertebrate photoreceptor, which unusually for a membrane protein is present in large quantities in the cell. It is clear that this structural information cannot necessarily be used to model other GPCRs. Progress in this field therefore relies on the recombinant production of large quantities of pure, functional membrane protein for biochemical and structural studies. The proposed project therefore unites the protein expression expertise of Roslyn Bill with the pharmacological expertise of David Poyner to investigate and develop eukaryotic expression systems to produce both full length and soluble forms of a unique GPCR, the CL-receptor, and its accessory protein, RAMP1. Roslyn Bill is known for her work on understanding the molecular basis of successful membrane protein overexpression in the eukaryotic hosts, Saccharomyces cerevisiae and Pichia pastoris. Her group has successfully produced a range of membrane proteins for crystallisation trials and subsequent structural analysis. The main theme of her work is in the development of novel expression hosts, for which she has submitted patent applications. Bill’s group will use these and other yeasts to produce CL and RAMP1 for structural and functional analysis. David Poyner has been working on the molecular pharmacology of CL/RAMP1, which is implicated in neurogenic inflammation, migraine and various cardiovascular disorders, for 15 years. The mode of action of RAMP1 is unknown but it and related RAMPs have now been shown to associate with around 10 GPCRs. Poyner’s group has successfully modelled both CL and RAMP1 and thus can suggest how they associate to produce functional receptors. The resulting proteins will be investigated structurally using NMR by Dr. B. Birdsall at the National Institute for Medical Research, and by X-ray crystallography by Dr Richard Neutze at Chalmers University, Gothenburg, Sweden. The project will be the catalyst for subsequent joint applications from Drs Bill and Poyner to the BBSRC and the Wellcome Trust, to investigate the structural basis of other GPCRs and their interactions with RAMPs, thereby laying the foundation for a deeper understanding of their pharmacology and facilitating novel drug design;