"PhD Studentship: The Role of O-glycosylation of the Stalk Region of the Immunoglobulin Receptor GPVI"

University of Birmingham - School of Medical Sciences

Background: Glycoprotein VI (GPVI) is the major signalling receptor for collagen on platelets and ligand binding to this receptor results in platelet activation. GPVI is a promising anti-thrombotic target due having major roles in thrombosis but limited roles in haemostasis, and GPVI inhibitors have shown promising results in clinical trials for preventing heart attack and stroke. GPVI is a single transmembrane receptor with two extracellular immunoglobulin domains, an O-glycosylated stalk, and a short intracellular tail. O-glycosylation at the stalk accounts for 50 % of the molecular weight of the receptor, but the exact locations of these glycosylation sites and their function are unknown. A recent antibody targeting the stalk region has been documented to have powerful anti-GPVI properties.

Aims: We believe the glycosylated stalk has potential to be a new region of the receptor to target with anti-platelet drugs. We aim to map the sites of O-glycosylation and determine their function on platelet signalling.

Objectives: In this project, students will map out the glycosylation sites through site directed mutagenesis. GPVI constructs lacking each glycosylation site will be tested in signalling assays to determine their effects on GPVI signalling. In addition, super resolution microscopy will be used to determine how glycosylation effects the distribution of GPVI on the cell membrane. Finally, nanobodies will be raised against the GPVI stalk region and their effects on GPVI expression and signalling will be monitored in transfected cell lines and on platelet cells.

Research environment: Students will be based in group of Dr Alexandre Slater and Prof Steve Watson within the Birmingham Platelet group at the University of Birmingham, Department of Cardiovascular Sciences. Dr Slater’s interests have focussed on studying platelet receptor structure whilst Prof Steve Watson’s interests are on platelet signalling. Both Dr Slater and Prof Watson are interested in developing novel inhibitors targeting GPVI.

Techniques and experimental approaches: This project will utilise a range of cell-based and biophysical techniques. A list of the major techniques are provided below:

• Mammalian cell culture
• Site directed mutagenesis
• Nanobody characterisation
• Size exclusion chromatography
• Super resolution microscopy
• Mass-spectrometry
• Platelet Biology

Outcomes: We will have a detailed understanding for the location and function of O-glycosylation sites within the stalk of GPVI. Nanobodies will be raised against key regions of the stalk and inhibitory nanobodies will be identified. The post potent nanobodies will be taken forward and developed as novel anti-platelet agents targeting thrombosis.

Person Specification: Applicants should have a strong background in either biomedicine, biochemistry or a related topic

How to apply: Informal enquiries should be directed to Dr Alexandre Slater (a.slater@bham.ac.uk) or Prof Steve Watson (s.p.watson@bham.ac.uk) who will form the supervisory team.

To make a formal application to the university, please follow this link, make an account, and submit an application via the university online admissions portal (via the above ‘Apply’ button). This link is unique to the MIBTP programme; please do not use any other link to apply to this project or your application may be rejected.

Applications should be directed to Dr Alexandre Slater (a.slater@bham.ac.uk) along with the following:

• A detailed CV.
• Names and addresses of two referees;
• A covering letter highlighting your research experience/capabilities;
• Copies of your degree certificates with transcripts;
• Evidence of your proficiency in the English language, if applicable.

For more details please visit: https://warwick.ac.uk/fac/cross_fac/mibtp/ or https://www.birmingham.ac.uk/about/college-of-life-and-environmental-sciences/midlands-integrative-biosciences-training-partnership