Toolkit of Approaches To Support Target-Focused Drug Discovery for Lysyl tRNA Synthetase

AUTHORS

Milne R , Wiedemar N , Corpas-Lopez V , Moynihan E , Wall RJ , Dawson A , Robinson DA , Shepherd SM , Smith RJ , Hallyburton I , Post JM , Dowers K , Torrie LS , Gilbert IH , Baragaña B , Patterson S , Wyllie S , . ACS infectious diseases. 2022 8 29; 8(9). 1962-1974

PMID: 36037410 [PubMed].

ABSTRACT

There is a pressing need for new medicines to prevent and treat malaria. Most antimalarial drug discovery is reliant upon phenotypic screening. However, with the development of improved target validation strategies, target-focused approaches are now being utilized. Here, we describe the development of a toolkit to support the therapeutic exploitation of a promising target, lysyl tRNA synthetase (KRS). The toolkit includes resistant mutants to probe resistance mechanisms and on-target engagement for specific chemotypes; a hybrid KRS protein capable of producing crystals suitable for ligand soaking, thus providing high-resolution structural information to guide compound optimization; chemical probes to facilitate pulldown studies aimed at revealing the full range of specifically interacting proteins and thermal proteome profiling (TPP); as well as streamlined isothermal TPP methods to provide unbiased confirmation of on-target engagement within a biologically relevant milieu. This combination of tools and methodologies acts as a template for the development of future target-enabling packages.

Tags: alumni publications 2022