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System-wide profiling of drug residence time by proteome integral solubility alteration assay

ABSTRACT: Most drugs used in the clinic and drug candidates target multiple proteins, and thus detailed characterization of their efficacy targets is required. While current methods rely on quantitative measurements at thermodynamic equilibrium, kinetic parameters such as the residence time of a drug on its target provide a better proxy for efficacy in vivo. Here, we present a new Residence Time Proteome Integral Solubility Alteration (ResT-PISA) assay which provides monitoring temporal protein solubility profiles after drug removal in either cell lysate or intact cells, quantifying the lifetime of the drug-target interaction. A compressed version of the assay measures the integral under the off-curve and enables the multiplexing of binding affinity proxy measurements with residence time assessment into a single proteomic analysis. We introduce a combined scoring system for three parametric dimensions to improve prioritization of targets. By providing complementary information to other characteristics of drug-target interaction, ResT-PISA approach can become a useful tool in drug development and precision medicine.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood

DISEASE(S): Acute Leukemia

SUBMITTER: Christian Beusch

LAB HEAD: Roman Zubarev

PROVIDER: PXD034351 | Pride | 2023-01-06

REPOSITORIES: Pride

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			Action	DRS
	200311_ResTime_LysateStaurosporine_repA_f1.raw	Raw
	200311_ResTime_LysateStaurosporine_repA_f10.raw	Raw
	200311_ResTime_LysateStaurosporine_repA_f11.raw	Raw
	200311_ResTime_LysateStaurosporine_repA_f12.raw	Raw
	200311_ResTime_LysateStaurosporine_repA_f13.raw	Raw

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