Studying Target–Engagement of Anti-Infectives by Solvent-Induced Protein Precipitation and Quantitative Mass Spectrometry DOI Creative Commons
Lorenzo Bizzarri,

Dominik Steinbrunn,

Thibaut Quennesson

et al.

ACS Infectious Diseases, Journal Year: 2024, Volume and Issue: 10(12), P. 4087 - 4102

Published: Nov. 20, 2024

Antimicrobial resistance (AMR) poses a serious threat to global health. The rapid emergence of contrasts with the slow pace antimicrobial development, emphasizing urgent need for innovative drug discovery approaches. This study addresses critical bottleneck in early development by introducing integral solvent-induced protein precipitation (iSPP) rapidly assess target–engagement lead compounds extracts pathogenic microorganisms under close-to-physiological conditions. iSPP measures change stability against presence ligands. method bacteria builds upon established SPP procedures and features optimized denaturation gradients minimized sample input amounts. effectiveness workflow was initially demonstrated through multidrug study. Using quantitative mass spectrometry (LC-MS/MS), we successfully identified known targets seven different antibiotics cell four AMR-related pathogens: three Gram-negative Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa Gram-positive bacterium Staphylococcus aureus. ultimately applied demonstrate derived from target-based discovery. We employed five small molecules targeting enzymes 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway─a promising focus anti-infective development. showcases adaptability efficiency identifying targets, advancing early-stage AMR.

Language: Английский

Introducing dysfunctional Protein-Protein Interactome (dfPPI) – A platform for systems-level protein-protein interaction (PPI) dysfunction investigation in disease DOI Creative Commons
Souparna Chakrabarty, Shujuan Wang, Tanaya Roychowdhury

et al.

Current Opinion in Structural Biology, Journal Year: 2024, Volume and Issue: 88, P. 102886 - 102886

Published: July 13, 2024

Protein-protein interactions (PPIs) play a crucial role in cellular function and disease manifestation, with dysfunctions PPI networks providing direct link between stressors phenotype. The dysfunctional Protein-Protein Interactome (dfPPI) platform, formerly known as epichaperomics, is newly developed chemoproteomic method aimed at detecting dynamic changes the systems level under stressor-induced perturbations within states. This review provides an overview of dfPPIs, emphasizing novel methodology, data analytics, applications research. dfPPI has cancer research, where it identifies integral to maintaining malignant phenotypes discovers strategies enhance efficacy current therapies. In neurodegenerative disorders, uncovers critical processes stressor-specific vulnerabilities. Challenges, including complexity potential for integration other omics datasets are discussed. platform potent tool dissecting biology by directly informing on holds promise advancing identification therapeutics.

Language: Английский

Citations

3
Studying Target–Engagement of Anti-Infectives by Solvent-Induced Protein Precipitation and Quantitative Mass Spectrometry DOI Creative Commons
Lorenzo Bizzarri,

Dominik Steinbrunn,

Thibaut Quennesson

et al.

ACS Infectious Diseases, Journal Year: 2024, Volume and Issue: 10(12), P. 4087 - 4102

Published: Nov. 20, 2024

Antimicrobial resistance (AMR) poses a serious threat to global health. The rapid emergence of contrasts with the slow pace antimicrobial development, emphasizing urgent need for innovative drug discovery approaches. This study addresses critical bottleneck in early development by introducing integral solvent-induced protein precipitation (iSPP) rapidly assess target–engagement lead compounds extracts pathogenic microorganisms under close-to-physiological conditions. iSPP measures change stability against presence ligands. method bacteria builds upon established SPP procedures and features optimized denaturation gradients minimized sample input amounts. effectiveness workflow was initially demonstrated through multidrug study. Using quantitative mass spectrometry (LC-MS/MS), we successfully identified known targets seven different antibiotics cell four AMR-related pathogens: three Gram-negative Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa Gram-positive bacterium Staphylococcus aureus. ultimately applied demonstrate derived from target-based discovery. We employed five small molecules targeting enzymes 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway─a promising focus anti-infective development. showcases adaptability efficiency identifying targets, advancing early-stage AMR.

Language: Английский

Citations

1