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

Rachel Milne; Natalie Wiedemar; Victoriano Corpas Lopez; Eoin Moynihan; Richard J. Wall; Alice Dawson; David A. Robinson; Sharon M. Shepherd; Robert J. Smith; Irene Hallyburton; John M. Post; Karen Dowers; Leah S. Torrie; Ian H. Gilbert; Beatriz Baragaña; Stephen Patterson; Susan Wyllie

doi:10.1021/acsinfecdis.2c00364

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

Rachel Milne, Natalie Wiedemar, Victoriano Corpas Lopez, Eoin Moynihan, Richard J. Wall, Alice Dawson, David A. Robinson, Sharon M. Shepherd, Robert J. Smith, Irene Hallyburton, John M. Post, Karen Dowers, Leah S. Torrie, Ian H. Gilbert, Beatriz Baragaña, Stephen Patterson, Susan Wyllie (Lead / Corresponding author)

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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 (PfKRS). 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.

Original language	English
Pages (from-to)	1962-1974
Number of pages	13
Journal	ACS Infectious Diseases
Volume	8
Issue number	9
Early online date	29 Aug 2022
DOIs	https://doi.org/10.1021/acsinfecdis.2c00364
Publication status	Published - 9 Sept 2022

Keywords

Plasmodium
lysyl tRNA synthetase
thermal proteome profiling (TPP)
isothermal TPP
chemical pulldown
anti-malarial drug discover
antimalarial drug discovery

ASJC Scopus subject areas

Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1021/acsinfecdis.2c00364Licence: CC BY

Final Published VersionFinal published version, 4.34 MBLicence: CC BY
Supporting InformationOther version, 945 KB

Cite this

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., Baragaña, B., Patterson, S., & Wyllie, S. (2022). Toolkit of Approaches To Support Target-Focused Drug Discovery for Plasmodium falciparum Lysyl tRNA Synthetase. ACS Infectious Diseases, 8(9), 1962-1974. https://doi.org/10.1021/acsinfecdis.2c00364

@article{05444c83a35a4fb4b2b41765ff1cdc74,

title = "Toolkit of Approaches To Support Target-Focused Drug Discovery for Plasmodium falciparum Lysyl tRNA Synthetase",

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 (PfKRS). 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.",

keywords = "Plasmodium, lysyl tRNA synthetase, thermal proteome profiling (TPP), isothermal TPP, chemical pulldown, anti-malarial drug discover, antimalarial drug discovery",

author = "Rachel Milne and Natalie Wiedemar and {Corpas Lopez}, Victoriano and Eoin Moynihan and Wall, {Richard J.} and Alice Dawson and Robinson, {David A.} and Shepherd, {Sharon M.} and Smith, {Robert J.} and Irene Hallyburton and Post, {John M.} and Karen Dowers and Torrie, {Leah S.} and Gilbert, {Ian H.} and Beatriz Baraga{\~n}a and Stephen Patterson and Susan Wyllie",

note = "Funding Information: This work was supported by the following funding from the Bill and Melinda Gates Foundation [OPP1193840], [OPP1032518], [OPP1032548] and [INV026519]. We would also like to acknowledge support from the Wellcome Trust in the form of an Innovations award (Mode of Action group, [218448/Z/19/Z]) and a centre award (The Wellcome Centre for Anti-Infectives Research, [203134/Z/16/Z]). We thank Diamond Light Source for beamtime (proposal mx10071) and the staff of beamline I24 for assistance with crystal testing and data collection. In addition, we would like the Scottish Blood Transfusion Centre (Ninewells Hospital, Dundee) for providing human erythrocytes for these studies. ",

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doi = "10.1021/acsinfecdis.2c00364",

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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 2022, 'Toolkit of Approaches To Support Target-Focused Drug Discovery for Plasmodium falciparum Lysyl tRNA Synthetase', ACS Infectious Diseases, vol. 8, no. 9, pp. 1962-1974. https://doi.org/10.1021/acsinfecdis.2c00364

TY - JOUR

T1 - Toolkit of Approaches To Support Target-Focused Drug Discovery for Plasmodium falciparum Lysyl tRNA Synthetase

AU - Milne, Rachel

AU - Wiedemar, Natalie

AU - Corpas Lopez, Victoriano

AU - Moynihan, Eoin

AU - Wall, Richard J.

AU - Dawson, Alice

AU - Robinson, David A.

AU - Shepherd, Sharon M.

AU - Smith, Robert J.

AU - Hallyburton, Irene

AU - Post, John M.

AU - Dowers, Karen

AU - Torrie, Leah S.

AU - Gilbert, Ian H.

AU - Baragaña, Beatriz

AU - Patterson, Stephen

AU - Wyllie, Susan

N1 - Funding Information: This work was supported by the following funding from the Bill and Melinda Gates Foundation [OPP1193840], [OPP1032518], [OPP1032548] and [INV026519]. We would also like to acknowledge support from the Wellcome Trust in the form of an Innovations award (Mode of Action group, [218448/Z/19/Z]) and a centre award (The Wellcome Centre for Anti-Infectives Research, [203134/Z/16/Z]). We thank Diamond Light Source for beamtime (proposal mx10071) and the staff of beamline I24 for assistance with crystal testing and data collection. In addition, we would like the Scottish Blood Transfusion Centre (Ninewells Hospital, Dundee) for providing human erythrocytes for these studies.

PY - 2022/9/9

Y1 - 2022/9/9

N2 - 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 (PfKRS). 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.

AB - 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 (PfKRS). 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.

KW - Plasmodium

KW - lysyl tRNA synthetase

KW - thermal proteome profiling (TPP)

KW - isothermal TPP

KW - chemical pulldown

KW - anti-malarial drug discover

KW - antimalarial drug discovery

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U2 - 10.1021/acsinfecdis.2c00364

DO - 10.1021/acsinfecdis.2c00364

M3 - Article

C2 - 36037410

SN - 2373-8227

VL - 8

SP - 1962

EP - 1974

JO - ACS Infectious Diseases

JF - ACS Infectious Diseases

IS - 9

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

A Platform for Drug Target Deconvolution and Exploitation

Wellcome Centre for Anti-Infectives Research

Structure-Guided Drug Discovery for Malaria and Tuberculosis (joint with Structural Genomics Consortium)

Cite this

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

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Projects

A Platform for Drug Target Deconvolution and Exploitation

Wellcome Centre for Anti-Infectives Research

Structure-Guided Drug Discovery for Malaria and Tuberculosis (joint with Structural Genomics Consortium)

Cite this