About the Project
G protein-coupled receptors, one of the most diverse and ubiquitous families of integral membrane proteins, play a pivotal role in many cellular signalling pathways. They are prime targets for the development of therapeutic agents designed to either block or activate these receptors. Although huge progress has been made in elucidating the structure of “Family A” GPCRs (e.g. β-adrengeric receptor structure led to the 2012 Nobel Prize in Chemistry), much less progress has been made with their cousins in “Family B”, despite their physiological importance and potential therapeutic potential for treating diabetes, migraine, pain, osteoporosis, etc.). We aim to elucidate the mechanism by which the Family B PTH2 receptor binds its ligand(s) and transduces a signal across the plasma membrane to initiate cell signalling. Techniques will include all general recombinant DNA work (subcloning, DNA preps, site-directed mutagenesis, etc), generation of recombinant baculovirus, over-expression and purification of receptor protein, transfection/culture of mammalian cells, stable cell line generation, radioligand binding assays, 384-well plate-based cell signalling assays, molecular modeling, crystallisation and crystallography.
Funding Notes
4 year BBSRC studentship, under the White Rose Mechanistic Biology DTP. The successful applicant will receive fees and stipend (c.£14,057). The PhD will start in Oct 2016. Applicants should have, or be expecting to receive, a 2.1 Hons degree in a relevant subject. EU candidates must have been resident in the UK for 3 years in order to receive full support. There are 2 stages to the application process: www.fbs.leeds.ac.uk/postgraduate/phdopportunities.php
References
Goldman:
Kellosalo, J., Kajander, T., Kogan, K., Pokharel, K. & Goldman, A. The Structure and Catalytic Cycle of a Sodium-Pumping Pyrophosphatase. Science 337, 473–476 (2012).
Tsai J-Y, Kellosalo J, Sun Y-J, Goldman A (2014) Proton/sodium pumping pyrophosphatases: the last of the primary ion pumps. Curr Opin Struct Biol, 27C: 38–47
Saarenpää, T. J., Jaakola, V.-P. & Goldman, A. Baculovirus mediated expression of GPCRs in insect cells. Methods Enzymol. 556, 185–218 (2015).
Meri, T., Amdahl, H., Lehtinen, M. J., Hyvärinen, S., McDowell, J. V., Bhattacharjee, A., Meri, S., Marconi, R., Goldman, A. & Jokiranta, T. S. Microbes bind complement inhibitor factor H via a common site. PLoS Pathog. 9, e1003308 (2013).
Donnelly:
Kumar A; Gopalswamy M; Wishart C; Henze M; Eschen-Lippold L; Donnelly D; Balbach J; Balbach J N-terminal phosphorylation of parathyroid hormone (PTH) abolishes its receptor activity ACS Chemical Biology 9 2465-2470, 2014
Weaver RE; Wigglesworth MJ; Wigglesworth MJ; Donnelly D A salt bridge between Arg-20 on parathyroid hormone (PTH) and Asp-137 on the PTH1 receptor is essential for full affinity Peptides 61 83-87, 2014
Mann RJ; Al-Sabah S; de Maturana RL; Sinfield JK; Donnelly D Functional coupling of Cys-226 and Cys-296 in the glucagon-like peptide-1 (GLP-1) receptor indicates a disulfide bond that is close to the activation pocket Peptides 31 2289-2293, 2010
Mann RJ; Nasr NE; Sinfield JK; Paci E; Donnelly D The major determinant of exendin-4/glucagon-like peptide 1 differential affinity at the rat glucagon-like peptide 1 receptor N-terminal domain is a hydrogen bond from SER-32 of exendin-4 Brit. J. Pharmacol 160 1973-1984, 2010
