| Autor: |
Nemmara VV; Department of Biochemistry and Molecular Pharmacology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States.; Program in Chemical Biology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States., Subramanian V; Department of Biochemistry and Molecular Pharmacology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States.; Program in Chemical Biology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States., Muth A; Department of Biochemistry and Molecular Pharmacology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States.; Program in Chemical Biology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States., Mondal S; Department of Biochemistry and Molecular Pharmacology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States.; Program in Chemical Biology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States., Salinger AJ; Department of Biochemistry and Molecular Pharmacology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States.; Program in Chemical Biology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States., Maurais AJ; Department of Chemistry , Boston College , Chestnut Hill , Massachusetts 02467 , United States., Tilvawala R; Department of Biochemistry and Molecular Pharmacology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States.; Program in Chemical Biology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States., Weerapana E; Department of Chemistry , Boston College , Chestnut Hill , Massachusetts 02467 , United States., Thompson PR; Department of Biochemistry and Molecular Pharmacology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States.; Program in Chemical Biology , UMass Medical School , 364 Plantation Street , Worcester , Massachusetts 01605 , United States. |
| Abstrakt: |
Citrullination is the post-translational hydrolysis of peptidyl-arginines to form peptidyl-citrulline, a reaction that is catalyzed by the protein arginine deiminases (PADs), a family of calcium-regulated enzymes. Aberrantly increased protein citrullination is associated with a slew of autoimmune diseases (e.g., rheumatoid arthritis (RA), multiple sclerosis, lupus, and ulcerative colitis) and certain cancers. Given the clear link between increased PAD activity and human disease, the PADs are therapeutically relevant targets. Herein, we report the development of next generation cell permeable and "clickable" probes (BB-Cl-Yne and BB-F-Yne) for covalent labeling of the PADs both in vitro and in cell-based systems. Using advanced chemoproteomic technologies, we also report the off targets of both BB-Cl-Yne and BB-F-Yne. The probes are highly specific for the PADs, with relatively few off targets, especially BB-F-Yne, suggesting the preferential use of the fluoroacetamidine warhead in next generation irreversible PAD inhibitors. Notably, these compounds can be used in a variety of modalities, including the identification of off targets of the parent compounds and as activity-based protein profiling probes in target engagement assays to demonstrate the efficacy of PAD inhibitors. |
