doi:10.7554/eLife.96343engMaity, RitwikZhang, XuepeiLiberati, Francesca RomanaScribani Rossi, ChiaraCutruzzolá, FrancescaRinaldo, SerenaGaetani, MassimilianoAínsa, José AntonioSancho, JavierMerging multi-omics with proteome integral solubility alteration unveils antibiotic mode of actionART-2024-140331Antimicrobial resistance is responsible for an alarming number of deaths, estimated at 5 million per year. To combat priority pathogens, like Helicobacter pylori, the development of novel therapies is of utmost importance. Understanding the molecular alterations induced by medications is critical for the design of multi-targeting treatments capable of eradicating the infection and mitigating its pathogenicity. However, the application of bulk omics approaches for unraveling drug molecular mechanisms of action is limited by their inability to discriminate between target-specific modifications and off-target effects. This study introduces a multi-omics method to overcome the existing limitation. For the first time, the Proteome Integral Solubility Alteration (PISA) assay is utilized in bacteria in the PISA-Express format to link proteome solubility with different and potentially immediate responses to drug treatment, enabling us the resolution to understand target-specific modifications and off-target effects. This study introduces a comprehensive method for understanding drug mechanisms and optimizing the development of multi-targeting antimicrobial therapies.2024http://zaguan.unizar.es/record/14546210.7554/eLife.96343http://zaguan.unizar.es/record/145462oai:zaguan.unizar.es:145462info:eu-repo/grantAgreement/ES/DGA/E45-23Rinfo:eu-repo/grantAgreement/EC/H2020/801586/EU/International Doctoral Programme for Talent Attraction to the Campus of International Excellence of the Ebro Valley/IberusTalentThis project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 801586-IberusTalentinfo:eu-repo/grantAgreement/ES/MICINN/PID2019-107293GB-I00info:eu-repo/grantAgreement/ES/MICINN/PID2022-141068NB-I00eLife 13 (2024), 21 pp.byhttp://creativecommons.org/licenses/by/3.0/es/info:eu-repo/semantics/openAccess