Peptidylarginine Deiminase and Citrullination: Potential Therapeutic Targets for Inflammatory Diseases

Byungki Jang; Sung Jae Shin

doi:10.4167/jbv.2013.43.3.159

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Jang and Shin: Peptidylarginine Deiminase and Citrullination: Potential Therapeutic Targets for Inflammatory Diseases

Review Article

J Bacteriol Virol 2013;43(3):159-167.

Published online: 9 February 2013

DOI: https://doi.org/10.4167/jbv.2013.43.3.159

Peptidylarginine Deiminase and Citrullination: Potential Therapeutic Targets for Inflammatory Diseases

Byungki Jang, Sung Jae Shin^*

Department of Microbiology and Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul, Korea

^*Corresponding author: Sung Jae Shin. Department of Microbiology and Institute of Immunology and Immunological Diseases, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752, Korea. Phone: +82-2-2228-1813, Fax: +82-2-392-9310, e-mail: sjshin@yuhs.ac

^** This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2013R1A1A2009822).

Received 12 July 201325 July 2013 Accepted 2 August 2013

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The multiple post-translational modifications of proteins display specific gain- or loss-of-function under normal and abnormal conditions. These modifications are precisely regulated by post-translational modification enzymes. The altered molecular status perturbs the pattern of gene expression and decides on a direction to signal transduction cascades as well as intrinsic properties of the proteins. Ultimately, it strictly maintains intracellular environment or results in disease manifestations. Recently, it has become that enzyme-dependent modification of arginine residue to citrulline exerts an important role in the induction of autoimmunity including rheumatoid arthritis, multiple sclerosis, and cancer. The modification of arginine residue to citrulline on proteins is called ‘citrullination’ or ‘deimination’ and is regulated by the calcium-dependent enzyme peptidylarginine deiminase (PAD). Now many effective PAD inhibitors (for example, Cl-amidine) have developed that ameliorates disease phenotypes. In this review, we discuss crucial roles of PAD enzyme and citrullination, the effectiveness of PAD inhibitors, and the implication in pathology.

Keywords: Peptidylarginine deiminase, Cl-amidine, Autoimmunity, Citrullination

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Figure 1.

Peptidylarginine deiminase (PAD) and citrullination. (A) The conversion of peptidylarginine to peptidylcitrulline is a posttranslational process driven by peptidylarginine deiminase (PAD) in a calcium (Ca²⁺) dependent manner. Ca²⁺ is an essential cofactor for the activation of PAD, effective Ca²⁺ concentration is approximately 100-fold higher than physiological cytosolic Ca²⁺ concentration. (B) Citrullination loss of target positive charge, alters protein structure and susceptibility of protease, changes natural functions, and become autoantigen. (C) Histone citrullination is involved in chromatin decondensation and formation of neutrophil extracellular traps. Citrullinated histones regulate gene transcription including p53- and estrogen receptor α-targeted. (D) PAD and citrullinated proteins are implicated in human diseases including cancer, rheumatoid arthritis, skin physiology, and neurodegeneration.

Figure 2.

Structures of Cl- and F-amidine, 2-Chloroacetamidine, and YW3-56. Each PAD inhibitors irreversibly inactivates PAD enzymes via the specific modification of Cys645, an active site residue that is critical for catalysis.

Table 1.

Peptidylarginine deiminase: protein expression and related diseases.

PAD type	Expression: protein	Disease	Target
PAD1	Epidermis, anagen hair follicle, keratinocytes, arrector pili muscles, sweat glands	Skin disorders	Keratins
PAD2	Epidermis, keratinocytes, arrector pili muscles, sweat glands, lung bronchoalveolar lavages (BAL), lymphocytes, macrophages, monocytes, neutrophils, oligodendrocytes, Schwann cells	Alzheimer's disease, Creutzfeldt-Jakob disease, multiple sclerosis, glaucoma, rheumatoid arthritis, Paget's disease Experimental autoimmune encephalomyelitis	Histone H3, vimentin, enolase, myelin basic protein, glial fibrillary acidic protein, tubulin
PAD3	Epidermis, anagen hair follicle, hair cuticle, keratinocytes, neutrophils, Schwann cells	Skin disorders	Keratins, S100A3
PAD4	Lung BAL, bone marrow, CD34+ cells, granulocytes, HL-60, lymphocytes, macrophages, monocytes	Apoptosis, cancer, multiple sclerosis, rheumatoid arthritis	Histone H3/H4, OKL38
PAD6	Embryo, oocyte	Rheumatoid arthritis	Unknown

Table 2.

Potent PAD inhibitors and in vivo test

Inhibitor	IC₅₀	In vitro and Animal models	Effects and Outcomes
		Collagen-induced arthritis	Reduced total synovial and serum citrullination Low histological score and complement C3 deposition Decreased IgG reactivity
Cl-amidine	5.9 ± 0.3 μM	New Zealand mixed 2328 model of murine lupus	Inhibition of NET formation Reduced circulating autoantibodies and complement levels Increased differentiation capacity of bone marrow endothelial progenitor cells Improved endothelium-dependent vasorelaxation Delayed time to arterial thrombosis
		DSS-induced colitis	Reduced the clinical signs and symptoms of colitis without toxic side effects Apoptosis of inflammatory cells
F-amidine	21.6 ± 2.1 μM	HL-60, MCF7, and HT-29 cancer cell lines	Decreased cell viability
2-CA	14.4 mM	ND4 mice model of multiple sclerosis	Dramatic attenuation of disease phenotype at any stage Reduced PAD activity and citrullination Suppression of T cell autoreactivity, clearing brain and spinal cord infiltrates
		Xenograft tumor model	Cancer growth inhibition
YW3-56	1∼5 μM	U2OS cell line	Activation of tumor suppression genes Inhibition of cancer cell growth Regulation of mTORC1 activity through SESN2/DDIT induction Regulation of autophagy flux by inhibiting autophagic vesicle breakdown by lysosomes

IC₅₀ is the concentration of the inhibitor that yields half-maximal activity. IC₅₀ values were determined by recombinant PAD enzymes in the presence of calcium and BAEE substrate (33∼35).

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