title: PAD2 Activity Monitored via a Fluorescent Substrate Analog.
abstract: The post-transitional modification of peptidyl arginine to citrulline by PAD2 can affect the inherent biophysical properties of the citrullinated protein. Furthermore, dysregulation of PAD2 activity has been implicated in a number of human diseases. Inhibition of these enzymes by small molecules can serve as essential probes in establishing a link to pathogenesis. Herein, we describe a profluorescent substrate analog that reports on the activity and the inhibition of PAD2 in a robust assay. Most noteworthy, we expect future drug discovery efforts based on PAD2 inhibition can be pursued via this assay.
authors: Chemistry Department, Lehigh University, Bethlehem, PA, 18015, USA.
authors: Mary J
authors: MJ
authors: Sabulski
authors: Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, PA, 16802, USA.
authors: Yanming
authors: Y
authors: Wang
authors: Chemistry Department, Lehigh University, Bethlehem, PA, 18015, USA.
authors: Marcos M
authors: MM
authors: Pires
mesh_terms: D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D001549:Benzamides / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002956:Citrulline / Q000378:metabolism; D003374:Coumarins / Q000378:metabolism; D057075:Enzyme Assays / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D011499:Protein Processing, Post-Translational; D000080064:Protein-Arginine Deiminase Type 2; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D013050:Spectrometry, Fluorescence / Q000379:methods; D014357:Trypsin / Q000378:metabolism
chemical_list: D001549:Benzamides; D003374:Coumarins; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; C000609001:YW4-03 compound; C041652:carbobenzoxyargininamide-7-yl-4-methylcoumarin; D002956:Citrulline; D001120:Arginine; D006867:Hydrolases; D014357:Trypsin; C000620352:PADI2 protein, human; C000629235:PADI4 protein, human; D000080064:Protein-Arginine Deiminase Type 2; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D002118:Calcium
keywords: PAD2; PAD4; citrullination; fluorescence assay; inhibitor