Utilizing thermal proteome profiling to identify the molecular targets of anti-leishmanial compounds

Victoriano Corpas Lopez; Susan Wyllie

doi:10.1016/j.xpro.2021.100704

Utilizing thermal proteome profiling to identify the molecular targets of anti-leishmanial compounds

Victoriano Corpas Lopez (Lead / Corresponding author), Susan Wyllie (Lead / Corresponding author)

Biological Chemistry and Drug Discovery

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

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Abstract

Here we detail our optimized protocol for the identification of drug targets in Leishmania donovani using thermal proteome profiling (TPP). This approach is based on the principle that binding of a drug to its protein target can significantly alter the thermal stability of that protein. By monitoring changes in the thermal stability of proteins within drug-treated and untreated cell lysates, using mass spectrometry combined with tandem-mass-tag labelling, putative targets of the drug can be identified in an unbiased manner.

Original language	English
Article number	100704
Number of pages	19
Journal	STAR Protocols
Volume	2
Issue number	3
Early online date	18 Aug 2021
DOIs	https://doi.org/10.1016/j.xpro.2021.100704
Publication status	Published - 17 Sept 2021

Keywords

Biophysics
Microbiology
Protein Biochemistry
Proteomics
Mass Spectrometry

Access to Document

10.1016/j.xpro.2021.100704Licence: CC BY-NC-ND

Final Published VersionFinal published version, 2.68 MBLicence: CC BY-NC-ND

1 Active
2 Finished

A Platform for Drug Target Deconvolution and Exploitation
Gilbert, I. (Investigator), Horn, D. (Investigator), Pawlowic, M. C. (Investigator), Wyatt, P. (Investigator) & Wyllie, S. (Investigator)
31/12/20 → 30/05/25
Project: Research
Wellcome Centre for Anti-Infectives Research
Cook, S. (Investigator), De Rycker, M. (Investigator), Fairlamb, A. (Investigator), Ferguson, M. (Investigator), Field, M. (Investigator), Gilbert, I. (Investigator), Gray, D. (Investigator), Horn, D. (Investigator), Pawlowic, M. C. (Investigator), Read, K. (Investigator), Wyatt, P. (Investigator) & Wyllie, S. (Investigator)
1/04/17 → 31/03/25
Project: Research
Chemical Biology: Leveraging Phenotypic Hits Against Kinetoplastids (Strategic Grant)
Fairlamb, A. (Investigator), Field, M. (Investigator), Gilbert, I. (Investigator), Gray, D. (Investigator), Horn, D. (Investigator) & Wyatt, P. (Investigator)
1/01/15 → 31/12/20
Project: Research

Fingerprints Proteomics Facility
Centre for Advanced Scientific Technologies
Facility/equipment: Facility

Cite this

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title = "Utilizing thermal proteome profiling to identify the molecular targets of anti-leishmanial compounds",

abstract = "Here we detail our optimized protocol for the identification of drug targets in Leishmania donovani using thermal proteome profiling (TPP). This approach is based on the principle that binding of a drug to its protein target can significantly alter the thermal stability of that protein. By monitoring changes in the thermal stability of proteins within drug-treated and untreated cell lysates, using mass spectrometry combined with tandem-mass-tag labelling, putative targets of the drug can be identified in an unbiased manner.",

keywords = "Biophysics, Microbiology, Protein Biochemistry, Proteomics, Mass Spectrometry",

author = "{Corpas Lopez}, Victoriano and Susan Wyllie",

note = "We would like to take this opportunity to thank the Fingerprints Proteomics Facility at the University of Dundee for their invaluable support throughout these studies. This work was supported by the following awards from the Wellcome Trust: Strategic Award [105021], Innovations Award [218448/Z/19/Z] and a Centre Award [203134/Z/16/Z].",

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AU - Corpas Lopez, Victoriano

AU - Wyllie, Susan

N1 - We would like to take this opportunity to thank the Fingerprints Proteomics Facility at the University of Dundee for their invaluable support throughout these studies. This work was supported by the following awards from the Wellcome Trust: Strategic Award [105021], Innovations Award [218448/Z/19/Z] and a Centre Award [203134/Z/16/Z].

PY - 2021/9/17

Y1 - 2021/9/17

N2 - Here we detail our optimized protocol for the identification of drug targets in Leishmania donovani using thermal proteome profiling (TPP). This approach is based on the principle that binding of a drug to its protein target can significantly alter the thermal stability of that protein. By monitoring changes in the thermal stability of proteins within drug-treated and untreated cell lysates, using mass spectrometry combined with tandem-mass-tag labelling, putative targets of the drug can be identified in an unbiased manner.

AB - Here we detail our optimized protocol for the identification of drug targets in Leishmania donovani using thermal proteome profiling (TPP). This approach is based on the principle that binding of a drug to its protein target can significantly alter the thermal stability of that protein. By monitoring changes in the thermal stability of proteins within drug-treated and untreated cell lysates, using mass spectrometry combined with tandem-mass-tag labelling, putative targets of the drug can be identified in an unbiased manner.

KW - Biophysics

KW - Microbiology

KW - Protein Biochemistry

KW - Proteomics

KW - Mass Spectrometry

U2 - 10.1016/j.xpro.2021.100704

DO - 10.1016/j.xpro.2021.100704

M3 - Article

C2 - 34467225

SN - 2666-1667

VL - 2

JO - STAR Protocols

JF - STAR Protocols

IS - 3

M1 - 100704

ER -

Utilizing thermal proteome profiling to identify the molecular targets of anti-leishmanial compounds

Abstract

Keywords

Access to Document

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Projects

A Platform for Drug Target Deconvolution and Exploitation

Wellcome Centre for Anti-Infectives Research

Chemical Biology: Leveraging Phenotypic Hits Against Kinetoplastids (Strategic Grant)

Equipment

Fingerprints Proteomics Facility

Cite this