Toolkit of Approaches To Support Target-Focused Drug Discovery for Plasmodium falciparum Lysyl tRNA Synthetase.
Milne, R., Wiedemar, N., Corpas-Lopez, V., Moynihan, E., Wall, R.J., Dawson, A., Robinson, D.A., Shepherd, S.M., Smith, R.J., Hallyburton, I., Post, J.M., Dowers, K., Torrie, L.S., Gilbert, I.H., Baragana, B., Patterson, S., Wyllie, S.(2022) ACS Infect Dis 8: 1962-1974
- PubMed: 36037410 
- DOI: https://doi.org/10.1021/acsinfecdis.2c00364
- Primary Citation of Related Structures:  
7ZOG - 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 ( Pf 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.
Organizational Affiliation: 
Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, U.K.