One-Pot Time-Induced Proteome Integral Solubility Alteration Assay for Automated and Sensitive Drug-Target Identification.

Autor:	Meng Z; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden.; Chemical Proteomics Unit, Science for Life Laboratory (SciLifeLab), 17165 Stockholm, Sweden.; Chemical Proteomics, Swedish National Infrastructure for Biological Mass Spectrometry (BioMS), 17177 Stockholm, Sweden., Saei AA; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 17177 Stockholm, Sweden., Lyu H; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden.; Biomotif AB, 18212 Danderyd, Sweden., Gaetani M; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden.; Chemical Proteomics Unit, Science for Life Laboratory (SciLifeLab), 17165 Stockholm, Sweden.; Chemical Proteomics, Swedish National Infrastructure for Biological Mass Spectrometry (BioMS), 17177 Stockholm, Sweden., Zubarev RA; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17177 Stockholm, Sweden.; Chemical Proteomics Unit, Science for Life Laboratory (SciLifeLab), 17165 Stockholm, Sweden.; Chemical Proteomics, Swedish National Infrastructure for Biological Mass Spectrometry (BioMS), 17177 Stockholm, Sweden.; Department of Pharmaceutical and Toxicological Chemistry, Medical Institute, Peoples' Friendship University of Russia named after Patrice Lumumba (RUDN University), 6 Miklukho-Maklaya St., Moscow 117198, Russian Federation.
Jazyk:	angličtina
Zdroj:	Analytical chemistry [Anal Chem] 2024 Dec 03; Vol. 96 (48), pp. 18917-18921. Date of Electronic Publication: 2024 Nov 20.
DOI:	10.1021/acs.analchem.4c05127
Abstrakt:	The proteome integral solubility alteration (PISA) assay is widely used for identifying drug targets, but it is labor-intensive and time-consuming and requires a substantial amount of biological sample. Aiming at enabling automation and greatly reducing the sample amount, we developed one-pot time-induced (OPTI)-PISA. Here, we demonstrate OPTI-PISA performance on identifying targets of multiple drugs in cell lysate and scaling down the sample amount to sub-microgram levels, making the PISA method suitable for NanoProteomics. OPTI-PISA can be implemented using only the standard equipment of a proteomics lab.
Databáze:	MEDLINE
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