Ubiquitylation impacts almost every biological process in the cell. Alterations in ubiquitin metabolism are related to the pathogenesis of cancer, neurodegenerative diseases, metabolic syndromes and inflammatory disorders. The enzymes involved in the ubiquitylation pathway (E1-E2-E3s ligases and Deubiquitylases) are the most promising next-generation targets for the development of specific therapeutics. Due to a lack of technologies that enable the exploration of this system in detail, there is still a gap between basic scientists, translational researchers and clinicians. In order to fill this gap and bring more ubiquitin targeting drugs to the market we have developed a high-throughput MALDI-TOF mass spectrometry assay. Through the use of 1,536 well plate format targets and liquid handling robots, very large numbers of compounds can be screened in an enzymatic in vitro assay. We have successfully applied this screen recently to identify specific Deubiquitylase (DUB) inhibitors (Ritorto et al, Nature Communications, 2014).
In this project, we will further develop high-throughput MALDI mass spectrometry with Bruker Daltonics. We will apply this exciting new technology to perform the first MALDI TOF drug discovery assay for E2/E3 ligases, which are also considered attractive drug targets. We will show proof-of-concept on MDM2 an E3 ubiquitin ligase with strong clinical relevance as it targets p53 for ubiquitin dependent degradation in the proteasome. The tumour suppressor p53 plays a pivotal role in protection from cancer development. Inhibitors of p53-MDM2 binding may offer an effective approach for cancer therapy. Small-molecules that disrupt the p53–MDM2 interaction have been identified and are currently in clinical evaluation. We are interested in applying our MALDI-TOF E2/E3 ligase assay to investigate the kinetic of p53 ubiquitylation and to assess the potency of already known and new potential inhibitors. Identified inhibitors will be further characterised using biophysical and cell biological tools and their activity will be tested in in vivo models. In this exciting project, we will show for the first time the high-throughput identification and characterisation of E2/E3 ligase inhibitor candidates by ultrafast MALDI TOF mass spectrometry.
Applicants should hold (or expect to obtain) a minimum upper-second honours degree (or equivalent) in chemistry, biology, biochemistry or a related field. In addition, candidates should be enthusiastic, able to work independently and thrive in a multi-disciplinary research environment. Strong interpersonal and communication skills are essential as is the ability to work as part of a team. Previous experience in mass spectrometry, analytical chemistry or drug discovery would be an advantage, but is not essential.
Our lab is equipped with seven state-of-the-art mass spectrometers including the newest ultra-fast RapiFlex MALDI TOF mass spectrometer. We offer hands-on training in a wide range of technologies including leading-edge proteomics techniques (such as mass spectrometry, chromatography, data analysis, statistics, bioinformatics) and drug discovery but also in cell biological and biochemical techniques.