Halogen Bonding Increases the Potency and Isozyme Selectivity of Protein Arginine Deiminase 1 Inhibitors

Mondal, Santanu; Gong, Xuefeng; Zhang, Xiaoqian; Salinger, Ari J.; Zheng, Li; Sen, Sudeshna; Weerapana, Eranthie; Zhang, Xuesen; Thompson, Paul R.

doi:10.1002/anie.201906334

Cited by 17 publications

(17 citation statements)

References 32 publications

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“…RFA labeling of WT and R374Cit PAD4. RFA labeling of PAD4 was carried out by following a protocol similar to that established for PAD1 and PAD2 10,11 . Briefly, PAD4 (100 nM final) was added to pre-warmed (10 min, 37°C) reaction mixture (100 mM Tris pH 7.4, 500 mM NaCl, 0 or 10 mM CaCl 2 , and 2 mM DTT in a final volume of 30 µL) containing RFA (200 nM final).…”

Section: Construction Of Plasmidsmentioning

confidence: 99%

“…Aberrant protein citrullination is a hallmark of multiple autoimmune disorders, including rheumatoid arthritis (RA), multiple sclerosis (MS), ulcerative colitis (UC), and lupus, as well as several neurodegenerative diseases and cancer. Of note, multiple pan-and isozyme-selective PAD inhibitors are known and these inhibitors show efficacy in animal models of RA, UC, MS, lupus, and sepsis 1,8,[10][11][12] . The contribution of protein hypercitrullination to the pathology of various diseases has been further established using the phenylglyoxal (PG)-based citrulline (Cit)specific probes, Rhodamine-PG (Rh-PG) and Biotin-PG 1,8 .…”

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confidence: 99%

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Site-specific incorporation of citrulline into proteins in mammalian cells

et al. 2021

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Citrullination is a post-translational modification (PTM) of arginine that is crucial for several physiological processes, including gene regulation and neutrophil extracellular trap formation. Despite recent advances, studies of protein citrullination remain challenging due to the difficulty of accessing proteins homogeneously citrullinated at a specific site. Herein, we report a technology that enables the site-specific incorporation of citrulline (Cit) into proteins in mammalian cells. This approach exploits an engineered E. coli-derived leucyl tRNA synthetase-tRNA pair that incorporates a photocaged-citrulline (SM60) into proteins in response to a nonsense codon. Subsequently, SM60 is readily converted to Cit with light in vitro and in living cells. To demonstrate the utility of the method, we biochemically characterize the effect of incorporating Cit at two known autocitrullination sites in Protein Arginine Deiminase 4 (PAD4, R372 and R374) and show that the R372Cit and R374Cit mutants are 181- and 9-fold less active than the wild-type enzyme. This technology possesses the potential to decipher the biology of citrullination.

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Section: Construction Of Plasmidsmentioning

confidence: 99%

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confidence: 99%

Site-specific incorporation of citrulline into proteins in mammalian cells

et al. 2021

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“…Other studies claim that PADs 4, 1, and 3 produce equimolar amounts of citrulline and ammonia [75]. PADs are calcium-dependent enzymes; the activity of PAD is highly regulated by calcium [74]. The requirement for multiple calcium ions is consistent with the structure of the calcium PAD 4 complex, showing that this enzyme binds up to 5 calcium ions at sites distal to the active site.…”

Section: Epigenetics and Posttranslational Mechanisms And Their Rementioning

confidence: 64%

“…PAD hydrolyzes the side chain of arginine to form citrulline. Citrullinated proteins are associated with autoimmune diseases that include RA and lupus and other pathologies [74]. In mammals, the PAD family consists of five members, namely PADs 1, 2, 3, 4, and 6, which exhibit tissue-specific expression patterns and varying in their subcellular location [75].…”

Section: Epigenetics and Posttranslational Mechanisms And Their Rementioning

confidence: 99%

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Epigenetic Mechanisms and Posttranslational Modifications in Systemic Lupus Erythematosus

Quiroz

Chavez-Estrada

Macias-Ochoa

et al. 2019

IJMS

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The complex physiology of eukaryotic cells is regulated through numerous mechanisms, including epigenetic changes and posttranslational modifications. The wide-ranging diversity of these mechanisms constitutes a way of dynamic regulation of the functionality of proteins, their activity, and their subcellular localization as well as modulation of the differential expression of genes in response to external and internal stimuli that allow an organism to respond or adapt to accordingly. However, alterations in these mechanisms have been evidenced in several autoimmune diseases, including systemic lupus erythematosus (SLE). The present review aims to provide an approach to the current knowledge of the implications of these mechanisms in SLE pathophysiology.

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Exploring Unconventional SAM Analogues To Build Cell‐Potent Bisubstrate Inhibitors for Nicotinamide N‐Methyltransferase

Iyamu¹,

Vilseck

Yadav

et al. 2022

Angewandte Chemie

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Nicotinamide N-methyltransferase (NNMT) methylates nicotinamide and has been associated with various diseases. Herein, we report the first cell-potent NNMT bisubstrate inhibitor II399, demonstrating a K i of 5.9 nM in a biochemical assay and a cellular IC 50 value of 1.9 μM. The inhibition mechanism and cocrystal structure confirmed II399 engages both the substrate and cofactor binding pockets. Computational modeling and binding data reveal a balancing act between enthalpic and entropic components that lead to II399's low nM binding affinity. Notably, II399 is 1 000-fold more selective for NNMT than closely related methyltransferases. We expect that II399 would serve as a valuable probe to elucidate NNMT biology. Furthermore, this strategy provides the first case of introducing unconventional SAM mimics, which can be adopted to develop cell-potent inhibitors for other SAM-dependent methyltransferases.

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Halogen Bonding Increases the Potency and Isozyme Selectivity of Protein Arginine Deiminase 1 Inhibitors

Abstract: Supportinginformation and the ORCID identification number(s) for the author(s) of this article can be found under: https://doi.

Cited by 17 publications

References 32 publications

Site-specific incorporation of citrulline into proteins in mammalian cells

Site-specific incorporation of citrulline into proteins in mammalian cells

Epigenetic Mechanisms and Posttranslational Modifications in Systemic Lupus Erythematosus

Exploring Unconventional SAM Analogues To Build Cell‐Potent Bisubstrate Inhibitors for Nicotinamide N‐Methyltransferase

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