The Computational Selection of Druggable Targets for the Development of Antifungals

Abstract

Fungal infections are a serious concern to the health and wellbeing of people worldwide. Up to 1.2 billion people a year suffer from fungal infections, although the majority of these infections are of the skin such as yeast infections, thrush, or athletes’ foot a considerable number are life-threatening. Between 1.5 and 2 million people a year die from invasive fungal infections. The infectious fungal species that opportunistically target those patients with a compromised immune system such as that resulting from from various cancer treatments, AIDS/HIV or organ transplant patients. The current drugs available are often ineffective due to fungal resistance or have serious side effects that can severely harm the patients. Research has shown a select number of proteins from 2 families of enzymes, Phosphatases and Kinases are essential to the growth and viability of multiple pathogenic fungi. In this study, I report an in-depth analysis, using computational tools, of some essential proteins and determine strategies to develop inhibitors that will target specific sites causing the death of the organism. I also report a complete protocol to express and purify one of the essential proteins, NimT. I demonstrate that NimT is an acid phosphatase and identify a number of inhibitors that halt the function of the protein dephosphorylation. I also take considerable steps to validate an additional site, alternative to that of the active site, as a potential drug target. In addition, we report a protocol that produces clean, soluble NimT protein that is suitable for structure determination crystallisation trials.

Date of Award	14 Oct 2022
Original language	English
Awarding Institution	The University of Manchester
Supervisor	Lydia Tabernero (Main Supervisor), Michael Bromley (Co Supervisor) & David Procter (Co Supervisor)