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Structure-based tailoring of the first coumarins-specific bergaptol O-methyltransferase to synthesize bergapten for depigmentation disorder treatment

Bergapten has long been used in combination with ultraviolet A irradiation to treat depigmentation disorder. However, extremely low bergapten contents in plants and difficulties in synthesizing bergapten have limited its application. Here, we developed an alternative bergapten-production method. We...

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Autores principales: Zhao, Yucheng, Wang, Nana, Wu, Huali, Zhou, Yuanze, Huang, Chuanlong, Luo, Jun, Zeng, Zhixiong, Kong, Lingyi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6812039/
https://www.ncbi.nlm.nih.gov/pubmed/31666994
http://dx.doi.org/10.1016/j.jare.2019.10.003
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author Zhao, Yucheng
Wang, Nana
Wu, Huali
Zhou, Yuanze
Huang, Chuanlong
Luo, Jun
Zeng, Zhixiong
Kong, Lingyi
author_facet Zhao, Yucheng
Wang, Nana
Wu, Huali
Zhou, Yuanze
Huang, Chuanlong
Luo, Jun
Zeng, Zhixiong
Kong, Lingyi
author_sort Zhao, Yucheng
collection PubMed
description Bergapten has long been used in combination with ultraviolet A irradiation to treat depigmentation disorder. However, extremely low bergapten contents in plants and difficulties in synthesizing bergapten have limited its application. Here, we developed an alternative bergapten-production method. We first determined the crystal structures of bergaptol O-methyltransferase from Peucedanum praeruptorum (PpBMT) and the ternary PpBMT–S-adenosyl-L-homocysteine (SAH)–bergaptol complex to identify key residues involved in bergaptol binding. Then, structure-based protein engineering was performed to obtain PpBMT mutants with improved catalytic activity towards bergaptol. Subsequently, a high-activity mutant was used to produce bergapten for pharmacological-activity analysis. Key PpBMT amino acids involved in bergaptol binding and substrate specificity were identified, such as Asp226, Asp246, Ser265, and Val320. Site-directed mutagenesis and biochemical analysis revealed that the V320I mutant efficiently transformed bergaptol to produce bergapten. Pharmacological-activity analysis indicated that bergapten positively affected hair pigmentation in mice and improved pigmentation levels in zebrafish embryos. This report provides the first description of the catalytic mechanism of coumarins-specific O-methyltransferase. The high-activity V320I mutant protein could be used in metabolic engineering to produce bergapten in order to treat depigmentation disorder. This structure–function study provides an alternative synthesis method and important advances for treating depigmentation disorders.
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spelling pubmed-68120392019-10-30 Structure-based tailoring of the first coumarins-specific bergaptol O-methyltransferase to synthesize bergapten for depigmentation disorder treatment Zhao, Yucheng Wang, Nana Wu, Huali Zhou, Yuanze Huang, Chuanlong Luo, Jun Zeng, Zhixiong Kong, Lingyi J Adv Res Original Article Bergapten has long been used in combination with ultraviolet A irradiation to treat depigmentation disorder. However, extremely low bergapten contents in plants and difficulties in synthesizing bergapten have limited its application. Here, we developed an alternative bergapten-production method. We first determined the crystal structures of bergaptol O-methyltransferase from Peucedanum praeruptorum (PpBMT) and the ternary PpBMT–S-adenosyl-L-homocysteine (SAH)–bergaptol complex to identify key residues involved in bergaptol binding. Then, structure-based protein engineering was performed to obtain PpBMT mutants with improved catalytic activity towards bergaptol. Subsequently, a high-activity mutant was used to produce bergapten for pharmacological-activity analysis. Key PpBMT amino acids involved in bergaptol binding and substrate specificity were identified, such as Asp226, Asp246, Ser265, and Val320. Site-directed mutagenesis and biochemical analysis revealed that the V320I mutant efficiently transformed bergaptol to produce bergapten. Pharmacological-activity analysis indicated that bergapten positively affected hair pigmentation in mice and improved pigmentation levels in zebrafish embryos. This report provides the first description of the catalytic mechanism of coumarins-specific O-methyltransferase. The high-activity V320I mutant protein could be used in metabolic engineering to produce bergapten in order to treat depigmentation disorder. This structure–function study provides an alternative synthesis method and important advances for treating depigmentation disorders. Elsevier 2019-10-10 /pmc/articles/PMC6812039/ /pubmed/31666994 http://dx.doi.org/10.1016/j.jare.2019.10.003 Text en © 2019 The Authors. Published by Elsevier B.V. on behalf of Cairo University. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Zhao, Yucheng
Wang, Nana
Wu, Huali
Zhou, Yuanze
Huang, Chuanlong
Luo, Jun
Zeng, Zhixiong
Kong, Lingyi
Structure-based tailoring of the first coumarins-specific bergaptol O-methyltransferase to synthesize bergapten for depigmentation disorder treatment
title Structure-based tailoring of the first coumarins-specific bergaptol O-methyltransferase to synthesize bergapten for depigmentation disorder treatment
title_full Structure-based tailoring of the first coumarins-specific bergaptol O-methyltransferase to synthesize bergapten for depigmentation disorder treatment
title_fullStr Structure-based tailoring of the first coumarins-specific bergaptol O-methyltransferase to synthesize bergapten for depigmentation disorder treatment
title_full_unstemmed Structure-based tailoring of the first coumarins-specific bergaptol O-methyltransferase to synthesize bergapten for depigmentation disorder treatment
title_short Structure-based tailoring of the first coumarins-specific bergaptol O-methyltransferase to synthesize bergapten for depigmentation disorder treatment
title_sort structure-based tailoring of the first coumarins-specific bergaptol o-methyltransferase to synthesize bergapten for depigmentation disorder treatment
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6812039/
https://www.ncbi.nlm.nih.gov/pubmed/31666994
http://dx.doi.org/10.1016/j.jare.2019.10.003
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