Understanding intrinsically disordered proteins and their potential as new drug targets through novel measurement capabilities

Research into the structure-function relationship of proteins has been a productive area of study over many decades, contributing to our understanding of biology and health and hence to the development of therapies. Intrinsically disordered proteins (IDPs) are a recently discovered class of proteins that are particularly important to study because of their overrepresentation in major diseases, including cancers and neurodegenerative diseases. Unlike most proteins that exist in a defined 3-dimensional structure, IDPs switch readily between multiple shapes. This renders them notoriously difficult to study. During this Fellowship novel methods will be developed to investigate IDPs, which will lead to deeper understanding of their behaviour, and will hence allow us to identify new drugs to manipulate them in disease states.
Nano-electrospray ionisation mass spectrometry (nESI-MS) is a powerful tool to probe protein dynamics, providing information on the range of shapes in which a protein exists. Furthermore, its low sample consumption, rapid data acquisition and low data processing positions MS as an effective tool in protein structure research. The hybrid technique of ion mobility-mass spectrometry (IMMS) provides further insights into the range of structural arrangements adopted by proteins and protein complexes, since here the ions are separated according to their size and their shape.
During the proposed research, state-of-the-art IMMS methods will be used to characterise specific IDPs that are involved in disease pathways. Proteins that are of timely significance will be chosen for these studies, and will include the disordered region of the androgen receptor that is heavily implicated in prostate cancer. The range of shapes that the IDPs exist in will be explored, and their interaction with other proteins that they're known to bind to in the cell will be described. If a small molecule has previously been identified as a potential drug, the influence that it has on the IDPs behaviour and its interaction with binding partners will be measured. Methods will be developed to test libraries of small molecules to find more potential drugs that affect the properties of the IDP in the appropriate manner. In-depth analysis of the protein-drug interactions will then be performed to assess their suitability as potential drug molecules.
Currently, IDP analysis is generally performed using techniques that require expensive instrumentation and specialist technological knowledge. Such approaches include nuclear magnetic resonance (NMR) and small angle scattering methods. This fellowship will enable analytical chemistry methods to be developed to create cost-effective, accessible systems for IDP characterisation by the scientific community. The applicability of various chromatographic methods, in which proteins are separated in solution according to properties such as their charge and shape, will be tested. Possibilities to develop low-cost, easy-to-use IMMS methods for protein characterisation will also be explored.
Improved care for our ageing society is currently an important research priority across many fields. This includes the development of novel therapeutics to keep people healthier for longer. The proposed research will help to address this need by developing tools for the scientific community to exploit an unexplored set of drug targets and, by using state-of-the-art technologies, to identify small molecules as drug leads for further testing. The proposed research is therefore likely to be highly beneficial for society.

Biotechnological and pharmaceutical companies will benefit from the development of innovative methods to exploit a new class of proteins as therapeutic targets, and identify drug leads that target them. IDPs were previously thought to be undruggable, in part due to the lack of methods with which to study them. The methods developed within the proposed research will help to overcome this notion. Additional benefits for the companies will include drug leads identified during the research. To maximise impact in this area, important drug targets will be used as test cases during the method development stages. This will include the disordered region of Androgen Receptor, a protein heavily implicated in castration-resistant prostate cancer. GlaxoSmithKline (Stevenage) hosts a large drug discovery department and they are likely to have an interest in methodologies that allow exploitation of a new class of drug targets. Existing links between Strathclyde and GSK will facilitate this cooperation.
The IDP community will benefit from knowledge about the conformational landscape of IDPs, which is impossible to visualise with other techniques. They will also benefit from the research that will be facilitated as a result of novel techniques for the community to measure IDPs, which will lead to an increased understanding, and extend the reach of the community.
The IMMS community will benefit from chromatographic methods that will serve as appropriate complementary techniques with which to validate gas-phase findings. Innovative methods for screening small molecule libraries for IDP inhibitors is likely to open up new areas of high-impact research, since it facilitates the possibility of identifying novel drugs for the treatment of important diseases.
Successful drug candidates identified during the research, or as a result of the novel methods developed during the Fellowship, will be of high benefit to society since it will allow better treatment of diseases. The need for new medicines has never been greater due to our aging population, and the medicines developed against cancer and neurodegenerative diseases will increase the standard of living in later life.
MS vendors will benefit from the research due to potential commercialisation of the innovative drug-screening methodologies. The methods will be developed on a Synapt G2S (Waters, UK). Existing links between the Fellow and Waters will provide the necessary means for maximum exploitation of the methods. The research could be of economic benefit therein, due to increased purchase of instrumentation, consumables and materials from Waters by users wishing to use the methods.
Research will be conducted to assess the suitability of Owlstone ultraFAIMS for studying native protein structure, with a specific focus on IDPs. The long-term aim of this is to develop an accessible, cost-effective standalone IMS device, in the absence of MS, for analysing protein structure and for drug screening purposes. Users will benefit from having an easy-to-use IMS device to analyse complex mixtures of protein structures, and Owlstone will benefit from research during the Fellowship since a concept and product for future commercialisation will be developed with potential economic impact.
I will train the PDRAs in the application of IMMS to IDPs. They will be trained in research-related and transferrable aspects of academic life, which will help them to fulfil their potential as future leaders. This will enable them to continue to make impact in the future.