Acefylline activates filaggrin deimination by peptidylarginine deiminases in the upper epidermis
•Screening of a large chemical library identified xanthines as PAD potential activators.•Deimination of human filaggrin was enhanced by caffeine, theobromine and acefylline.•Topical application of acefylline on reconstructed human epidermis improved deimination.•Acefylline is the first identified ac...
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| Published in: | Journal of dermatological science Vol. 81; no. 2; pp. 101 - 106 |
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| Main Authors: | , , , , , , , , , , |
| Format: | Journal Article |
| Language: | English |
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Elsevier Ireland Ltd
01.02.2016
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| ISSN: | 0923-1811, 1873-569X, 1873-569X |
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| Abstract | •Screening of a large chemical library identified xanthines as PAD potential activators.•Deimination of human filaggrin was enhanced by caffeine, theobromine and acefylline.•Topical application of acefylline on reconstructed human epidermis improved deimination.•Acefylline is the first identified activator of PAD1 and PAD3.
Peptidylarginine deiminases (PADs) catalyze deimination (or citrullination), a calcium-dependent post-translational modification involved in several physiological processes and human diseases, such as rheumatoid arthritis and cancer. Deimination of filaggrin (FLG) by PAD1 and PAD3 during the last steps of keratinocyte differentiation is a crucial event for the epidermis function and homeostasis. This allows the complete degradation of FLG, leading to the production of free amino acids and their derivatives that are essential for epidermal photoprotection and moisturizing of the stratum corneum.
To increase the flux of this catabolic pathway, we searched for activators of PADs.
A large chemical library was screened first in silico and then by using an automated assay based on an indirect colorimetric measurement of recombinant human PAD activity. Potential activators were then confirmed using a recombinant human FLG as a substrate, and secondly after topical application at the surface of three-dimensional reconstructed human epidermis.
The data obtained after the library screening pointed to xanthine derivatives as potential PAD activators. Among seven xanthine derivatives tested at 50–300μM, caffeine, theobromine and acefylline proved to be the most potent enhancers of in vitro deimination of FLG by PAD1 and PAD3. After topical application of a gel formulation containing 3% acefylline at the surface of reconstructed epidermis, immunoblotting analysis showed an increase in the total amount of deiminated proteins, and confocal microscopy showed an enhanced deimination in the stratum corneum. This demonstrated the activation of PADs in living cells.
As a PAD activator, acefylline will be useful to study the role of deimination and could be proposed to increase or correct the hydration of the cornified layers of the epidermis. |
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| AbstractList | Peptidylarginine deiminases (PADs) catalyze deimination (or citrullination), a calcium-dependent post-translational modification involved in several physiological processes and human diseases, such as rheumatoid arthritis and cancer. Deimination of filaggrin (FLG) by PAD1 and PAD3 during the last steps of keratinocyte differentiation is a crucial event for the epidermis function and homeostasis. This allows the complete degradation of FLG, leading to the production of free amino acids and their derivatives that are essential for epidermal photoprotection and moisturizing of the stratum corneum.BACKGROUNDPeptidylarginine deiminases (PADs) catalyze deimination (or citrullination), a calcium-dependent post-translational modification involved in several physiological processes and human diseases, such as rheumatoid arthritis and cancer. Deimination of filaggrin (FLG) by PAD1 and PAD3 during the last steps of keratinocyte differentiation is a crucial event for the epidermis function and homeostasis. This allows the complete degradation of FLG, leading to the production of free amino acids and their derivatives that are essential for epidermal photoprotection and moisturizing of the stratum corneum.To increase the flux of this catabolic pathway, we searched for activators of PADs.OBJECTIVETo increase the flux of this catabolic pathway, we searched for activators of PADs.A large chemical library was screened first in silico and then by using an automated assay based on an indirect colorimetric measurement of recombinant human PAD activity. Potential activators were then confirmed using a recombinant human FLG as a substrate, and secondly after topical application at the surface of three-dimensional reconstructed human epidermis.METHODSA large chemical library was screened first in silico and then by using an automated assay based on an indirect colorimetric measurement of recombinant human PAD activity. Potential activators were then confirmed using a recombinant human FLG as a substrate, and secondly after topical application at the surface of three-dimensional reconstructed human epidermis.The data obtained after the library screening pointed to xanthine derivatives as potential PAD activators. Among seven xanthine derivatives tested at 50-300μM, caffeine, theobromine and acefylline proved to be the most potent enhancers of in vitro deimination of FLG by PAD1 and PAD3. After topical application of a gel formulation containing 3% acefylline at the surface of reconstructed epidermis, immunoblotting analysis showed an increase in the total amount of deiminated proteins, and confocal microscopy showed an enhanced deimination in the stratum corneum. This demonstrated the activation of PADs in living cells.RESULTSThe data obtained after the library screening pointed to xanthine derivatives as potential PAD activators. Among seven xanthine derivatives tested at 50-300μM, caffeine, theobromine and acefylline proved to be the most potent enhancers of in vitro deimination of FLG by PAD1 and PAD3. After topical application of a gel formulation containing 3% acefylline at the surface of reconstructed epidermis, immunoblotting analysis showed an increase in the total amount of deiminated proteins, and confocal microscopy showed an enhanced deimination in the stratum corneum. This demonstrated the activation of PADs in living cells.As a PAD activator, acefylline will be useful to study the role of deimination and could be proposed to increase or correct the hydration of the cornified layers of the epidermis.CONCLUSIONAs a PAD activator, acefylline will be useful to study the role of deimination and could be proposed to increase or correct the hydration of the cornified layers of the epidermis. •Screening of a large chemical library identified xanthines as PAD potential activators.•Deimination of human filaggrin was enhanced by caffeine, theobromine and acefylline.•Topical application of acefylline on reconstructed human epidermis improved deimination.•Acefylline is the first identified activator of PAD1 and PAD3. Peptidylarginine deiminases (PADs) catalyze deimination (or citrullination), a calcium-dependent post-translational modification involved in several physiological processes and human diseases, such as rheumatoid arthritis and cancer. Deimination of filaggrin (FLG) by PAD1 and PAD3 during the last steps of keratinocyte differentiation is a crucial event for the epidermis function and homeostasis. This allows the complete degradation of FLG, leading to the production of free amino acids and their derivatives that are essential for epidermal photoprotection and moisturizing of the stratum corneum. To increase the flux of this catabolic pathway, we searched for activators of PADs. A large chemical library was screened first in silico and then by using an automated assay based on an indirect colorimetric measurement of recombinant human PAD activity. Potential activators were then confirmed using a recombinant human FLG as a substrate, and secondly after topical application at the surface of three-dimensional reconstructed human epidermis. The data obtained after the library screening pointed to xanthine derivatives as potential PAD activators. Among seven xanthine derivatives tested at 50–300μM, caffeine, theobromine and acefylline proved to be the most potent enhancers of in vitro deimination of FLG by PAD1 and PAD3. After topical application of a gel formulation containing 3% acefylline at the surface of reconstructed epidermis, immunoblotting analysis showed an increase in the total amount of deiminated proteins, and confocal microscopy showed an enhanced deimination in the stratum corneum. This demonstrated the activation of PADs in living cells. As a PAD activator, acefylline will be useful to study the role of deimination and could be proposed to increase or correct the hydration of the cornified layers of the epidermis. Background: Peptidylarginine deiminases (PADs) catalyze deimination (or citrullination), a calcium-dependent post-translational modification involved in several physiological processes and human diseases, such as rheumatoid arthritis and cancer. Deimination of filaggrin (FLG) by PAD1 and PAD3 during the last steps of keratinocyte differentiation is a crucial event for the epidermis function and homeostasis. This allows the complete degradation of FLG, leading to the production of free amino acids and their derivatives that are essential for epidermal photoprotection and moisturizing of the stratum corneum. Objective: To increase the flux of this catabolic pathway, we searched for activators of PADs. Methods: A large chemical library was screened first in silico and then by using an automated assay based on an indirect colorimetric measurement of recombinant human PAD activity. Potential activators were then confirmed using a recombinant human FLG as a substrate, and secondly after topical application at the surface of three-dimensional reconstructed human epidermis. Results: The data obtained after the library screening pointed to xanthine derivatives as potential PAD activators. Among seven xanthine derivatives tested at 50-300μM, caffeine, theobromine and acefylline proved to be the most potent enhancers of in vitro deimination of FLG by PAD1 and PAD3. After topical application of a gel formulation containing 3% acefylline at the surface of reconstructed epidermis, immunoblotting analysis showed an increase in the total amount of deiminated proteins, and confocal microscopy showed an enhanced deimination in the stratum corneum. This demonstrated the activation of PADs in living cells. Conclusion: As a PAD activator, acefylline will be useful to study the role of deimination and could be proposed to increase or correct the hydration of the cornified layers of the epidermis. Peptidylarginine deiminases (PADs) catalyze deimination (or citrullination), a calcium-dependent post-translational modification involved in several physiological processes and human diseases, such as rheumatoid arthritis and cancer. Deimination of filaggrin (FLG) by PAD1 and PAD3 during the last steps of keratinocyte differentiation is a crucial event for the epidermis function and homeostasis. This allows the complete degradation of FLG, leading to the production of free amino acids and their derivatives that are essential for epidermal photoprotection and moisturizing of the stratum corneum. To increase the flux of this catabolic pathway, we searched for activators of PADs. A large chemical library was screened first in silico and then by using an automated assay based on an indirect colorimetric measurement of recombinant human PAD activity. Potential activators were then confirmed using a recombinant human FLG as a substrate, and secondly after topical application at the surface of three-dimensional reconstructed human epidermis. The data obtained after the library screening pointed to xanthine derivatives as potential PAD activators. Among seven xanthine derivatives tested at 50-300μM, caffeine, theobromine and acefylline proved to be the most potent enhancers of in vitro deimination of FLG by PAD1 and PAD3. After topical application of a gel formulation containing 3% acefylline at the surface of reconstructed epidermis, immunoblotting analysis showed an increase in the total amount of deiminated proteins, and confocal microscopy showed an enhanced deimination in the stratum corneum. This demonstrated the activation of PADs in living cells. As a PAD activator, acefylline will be useful to study the role of deimination and could be proposed to increase or correct the hydration of the cornified layers of the epidermis. Highlights • Screening of a large chemical library identified xanthines as PAD potential activators. • Deimination of human filaggrin was enhanced by caffeine, theobromine and acefylline. • Topical application of acefylline on reconstructed human epidermis improved deimination. • Acefylline is the first identified activator of PAD1 and PAD3. |
| Author | Galliano, Marie-Florence Vidaluc, Jean-Louis Serre, Guy Daunes-Marion, Sylvie Duplan, Hélène Méchin, Marie-Claire Takahara, Hidenari Bessou-Touya, Sandrine Simon, Michel Cau, Laura Poigny, Stéphane |
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| Keywords | FLG Post-translational modification PAD Epidermal barrier LUT Skin Xanthine Filaggrin lookup table peptidylarginine deiminase filaggrin |
| Language | English |
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| Snippet | •Screening of a large chemical library identified xanthines as PAD potential activators.•Deimination of human filaggrin was enhanced by caffeine, theobromine... Highlights • Screening of a large chemical library identified xanthines as PAD potential activators. • Deimination of human filaggrin was enhanced by caffeine,... Peptidylarginine deiminases (PADs) catalyze deimination (or citrullination), a calcium-dependent post-translational modification involved in several... Background: Peptidylarginine deiminases (PADs) catalyze deimination (or citrullination), a calcium-dependent post-translational modification involved in... |
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| SubjectTerms | Administration, Cutaneous Cells, Cultured Computer Simulation Dermatology Dose-Response Relationship, Drug Enzyme Activation Enzyme Activators - administration & dosage Enzyme Activators - chemistry Enzyme Activators - pharmacology Epidermal barrier Epidermal Cells Epidermis / drug effects Epidermis - cytology Epidermis - drug effects Epidermis - enzymology Filaggrin Human health and pathology Humans Humans Hydrolases / metabolism Hydrolases - metabolism Infectious diseases Intermediate Filament Proteins - chemistry Intermediate Filament Proteins - metabolism Keratinocytes - drug effects Keratinocytes - enzymology Life Sciences Models, Molecular Post-translational modification Protein Conformation Protein Processing, Post-Translational Protein-Arginine Deiminases Skin Small Molecule Libraries Structure-Activity Relationship Theophylline - administration & dosage Theophylline - analogs & derivatives Theophylline - chemistry Theophylline - pharmacology Xanthine |
| Title | Acefylline activates filaggrin deimination by peptidylarginine deiminases in the upper epidermis |
| URI | https://www.clinicalkey.com/#!/content/1-s2.0-S0923181115300785 https://www.clinicalkey.es/playcontent/1-s2.0-S0923181115300785 https://dx.doi.org/10.1016/j.jdermsci.2015.11.006 https://www.ncbi.nlm.nih.gov/pubmed/26616205 https://www.proquest.com/docview/1760922247 https://ut3-toulouseinp.hal.science/hal-03159207 |
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