UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology; Program in Chemical Biology; Thompson Lab

Publication Date


Document Type

Article Postprint


Biochemistry | Enzymes and Coenzymes | Medicinal-Pharmaceutical Chemistry | Therapeutics


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.


protein arginine deiminases, probes

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This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Chemical Biology, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see

DOI of Published Version



ACS Chem Biol. 2018 Mar 16;13(3):712-722. doi: 10.1021/acschembio.7b00957. Epub 2018 Feb 1. Link to article on publisher's website

Journal/Book/Conference Title

ACS Chemical Biology

Related Resources

Link to article in PubMed

PubMed ID


Nemmara et al 2017 BB_Fyne paper Supporting Information ver 18.pdf (916 kB)
Supporting Information

Figure-1-revised.tif (716 kB)
Figure 1

Figure-2 Revised.tif (1321 kB)
Figure 2

Figure-3.tif (638 kB)
Figure 3

Scheme-1.tif (303 kB)
Scheme 1. Synthesis of BB-F-Yne and BB-Cl-Yne

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Figure 4