The aim of this project is to engineer subunit-specific anti-GluN1/2/A/B polypeptide affinity reagents and to characterise their pharmacological specificity and functional activity on the recombinant NMDARs expressed in heterologous expression systems and on native NMDARs in hippocampal slice preparations.
N-methyl-D-aspartate receptors (NMDARs) play fundamental roles in central synaptic transmission, synaptic plasticity, pain perception, neuronal development and are implicated in many pathological changes and age-related neurodegenerative diseases. NMDARs subunit composition, allosteric regulation by ligands and modulators and the time course of Ca2+ rise may yield either long-term potentiation (LTP) or long-term depression (LTD) of excitatory synaptic transmission. However distinguishing and selective modulation of the native NMDARs pharmacologically remains problematical, due to the lack of subunit-specific ligands.
The existing dogma and the current approach to developing highly specific polypeptide based ligands or other affinity reagents (such as antibodies, antibody mimics, their fragments and aptamers against proteins) assumes and aims to achieve a high degree of complementarity at the interface between such a polypeptide or an affinity reagent and the surface of the target molecule. Such an interface is stabilised through a combination of hydrogen bonds (with or without additional water molecules), ionic and van der Waals interactions, and through spatial complementarity between the two molecules.
Artibody approach to the design of protein and sequence-specific peptide based affinity reagents, recently developed at RHUL, relies on the domain and loop-swapping, allowing us to replace labile structural elements of a protein by the same or similar elements of the engineered Artibody polypeptides. Because Artibody molecules are based on the target protein sequence, this approach should enable us to rationally design short peptide-based ligands for a given protein target.
This project is a collaboration between two groups at the School of Biological Sciences (Dr Soloviev and Dr Chen). Informal enquiries should be addressed to [email protected] and [email protected].
A successful applicant is expected to have first class, 2.i or equivalent degree from a recognised University and relevant experience in one of the following: Protein engineering and protein self-assembly, Biochemistry, Expression of recombinant receptors in heterologous expression systems (Xenopus oocytes and mammalian cell lines), Pharmacological and functional assessment of receptor function (two-electrode voltage clamp electrophysiology, whole cell patch clamp, site-directed mutagenesis and structure-function studies) and an interest in drug discovery, BioPharmacology, Medicinal Chemistry, Drug Metabolism & Pharmacokinetics