Development and use of clickable activity based protein profiling agents for protein arginine deiminase 4.
The protein arginine deiminases (PADs), which catalyze the hydrolysis of peptidyl-arginine to form peptidyl-citrulline, are potential targets for the development of a rheumatoid arthritis (RA) therapeutic, as well as other human diseases including colitis and cancer. Additionally, these enzymes, and in particular PAD4, appear to play important roles in a variety of cell signaling pathways including apoptosis, differentiation, and transcriptional regulation. To better understand the factors that regulate in vivo PAD4 activity, we set out to design and synthesize a series of activity-based protein profiling (ABPP) reagents that target this enzyme. Herein we describe the design, synthesis, and evaluation of six ABPPs including (i) FITC-conjugated F-amidine (FFA1 and 2) and Cl-amidine (FCA1 and 2), and (ii) biotin-conjugated F-amidine (BFA) and Cl-amidine (BCA). We further demonstrate the utility of these probes for labeling PAD4 in cells, as well as for isolating PAD4 and PAD4 binding proteins. These probes will undoubtedly prove to be powerful tools that can be used to dissect the factors controlling the dynamics of PAD4 expression, activity, and function.
The Department of Chemistry and Biochemistry, University of South Carolina, Columbia, South Carolina 29208, USA.
Jessica L
JL
Slack
Corey P
CP
Causey
Yuan
Y
Luo
Paul R
PR
Thompson
D000578:Amidines / Q000494:pharmacology*; D001710:Biotin / Q000031:analogs & derivatives; D004789:Enzyme Activation; D005456:Fluorescent Dyes; D006801:Humans; D006867:Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D057230:Limit of Detection; D040081:Protein Array Analysis; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D014230:Triazoles / Q000737:chemistry
D000578:Amidines; D005456:Fluorescent Dyes; D014230:Triazoles; D001710:Biotin; D006867:Hydrolases; C000629235:PADI4 protein, human; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases
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