Mechanism of action and inhibition of dehydrosqualene synthase

Fu Yang Lin, Chia I. Liu, Yi Liang Liu, Yonghui Zhang, Ke Wang, Wen Yih Jeng, Tzu Ping Ko, Rong Cao, Andrew H J Wang, Eric Oldfield

研究成果: 雜誌貢獻文章

42 引文 (Scopus)

摘要

"Head-to-head" terpene synthases catalyze the first committed steps in sterol and carotenoid biosynthesis: the condensation of two isoprenoid diphosphates to form cyclopropylcarbinyl diphosphates, followed by ring opening. Here, we report the structures of Staphylococcus aureus dehydrosqualene synthase (CrtM) complexed with its reaction intermediate, presqualene diphosphate (PSPP), the dehydrosqualene (DHS) product, as well as a series of inhibitors. The results indicate that, on initial diphosphate loss, the primary carbocation so formed bends down into the interior of the protein to react with C2,3 double bond in the prenyl acceptor to form PSPP, with the lower two-thirds of both PSPP chains occupying essentially the same positions as found in the two farnesyl chains in the substrates. The second-half reaction is then initiated by the PSPP diphosphate returning back to the Mg2+ cluster for ionization, with the resultant DHS so formed being trapped in a surface pocket. This mechanism is supported by the observation that cationic inhibitors (of interest as antiinfectives) bind with their positive charge located in the same region as the cyclopropyl carbinyl group; that S-thiolo-diphosphates only inhibit when in the allylic site; activity results on 11 mutants show that both DXXXD conserved domains are essential for PSPP ionization; and the observation that head-to-tail isoprenoid synthases as well as terpene cyclases have ionization and alkene-donor sites which spatially overlap those found in CrtM.
原文英語
頁(從 - 到)21337-21342
頁數6
期刊Proceedings of the National Academy of Sciences of the United States of America
107
發行號50
DOIs
出版狀態已發佈 - 十二月 14 2010
對外發佈Yes

指紋

Diphosphates
Terpenes
Alkenes
Sterols
Carotenoids
Staphylococcus aureus
presqualene pyrophosphate
dehydrosqualene
Proteins

ASJC Scopus subject areas

  • General

引用此文

Mechanism of action and inhibition of dehydrosqualene synthase. / Lin, Fu Yang; Liu, Chia I.; Liu, Yi Liang; Zhang, Yonghui; Wang, Ke; Jeng, Wen Yih; Ko, Tzu Ping; Cao, Rong; Wang, Andrew H J; Oldfield, Eric.

於: Proceedings of the National Academy of Sciences of the United States of America, 卷 107, 編號 50, 14.12.2010, p. 21337-21342.

研究成果: 雜誌貢獻文章

Lin, FY, Liu, CI, Liu, YL, Zhang, Y, Wang, K, Jeng, WY, Ko, TP, Cao, R, Wang, AHJ & Oldfield, E 2010, 'Mechanism of action and inhibition of dehydrosqualene synthase', Proceedings of the National Academy of Sciences of the United States of America, 卷 107, 編號 50, 頁 21337-21342. https://doi.org/10.1073/pnas.1010907107
Lin, Fu Yang ; Liu, Chia I. ; Liu, Yi Liang ; Zhang, Yonghui ; Wang, Ke ; Jeng, Wen Yih ; Ko, Tzu Ping ; Cao, Rong ; Wang, Andrew H J ; Oldfield, Eric. / Mechanism of action and inhibition of dehydrosqualene synthase. 於: Proceedings of the National Academy of Sciences of the United States of America. 2010 ; 卷 107, 編號 50. 頁 21337-21342.
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abstract = "{"}Head-to-head{"} terpene synthases catalyze the first committed steps in sterol and carotenoid biosynthesis: the condensation of two isoprenoid diphosphates to form cyclopropylcarbinyl diphosphates, followed by ring opening. Here, we report the structures of Staphylococcus aureus dehydrosqualene synthase (CrtM) complexed with its reaction intermediate, presqualene diphosphate (PSPP), the dehydrosqualene (DHS) product, as well as a series of inhibitors. The results indicate that, on initial diphosphate loss, the primary carbocation so formed bends down into the interior of the protein to react with C2,3 double bond in the prenyl acceptor to form PSPP, with the lower two-thirds of both PSPP chains occupying essentially the same positions as found in the two farnesyl chains in the substrates. The second-half reaction is then initiated by the PSPP diphosphate returning back to the Mg2+ cluster for ionization, with the resultant DHS so formed being trapped in a surface pocket. This mechanism is supported by the observation that cationic inhibitors (of interest as antiinfectives) bind with their positive charge located in the same region as the cyclopropyl carbinyl group; that S-thiolo-diphosphates only inhibit when in the allylic site; activity results on 11 mutants show that both DXXXD conserved domains are essential for PSPP ionization; and the observation that head-to-tail isoprenoid synthases as well as terpene cyclases have ionization and alkene-donor sites which spatially overlap those found in CrtM.",
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T1 - Mechanism of action and inhibition of dehydrosqualene synthase

AU - Lin, Fu Yang

AU - Liu, Chia I.

AU - Liu, Yi Liang

AU - Zhang, Yonghui

AU - Wang, Ke

AU - Jeng, Wen Yih

AU - Ko, Tzu Ping

AU - Cao, Rong

AU - Wang, Andrew H J

AU - Oldfield, Eric

PY - 2010/12/14

Y1 - 2010/12/14

N2 - "Head-to-head" terpene synthases catalyze the first committed steps in sterol and carotenoid biosynthesis: the condensation of two isoprenoid diphosphates to form cyclopropylcarbinyl diphosphates, followed by ring opening. Here, we report the structures of Staphylococcus aureus dehydrosqualene synthase (CrtM) complexed with its reaction intermediate, presqualene diphosphate (PSPP), the dehydrosqualene (DHS) product, as well as a series of inhibitors. The results indicate that, on initial diphosphate loss, the primary carbocation so formed bends down into the interior of the protein to react with C2,3 double bond in the prenyl acceptor to form PSPP, with the lower two-thirds of both PSPP chains occupying essentially the same positions as found in the two farnesyl chains in the substrates. The second-half reaction is then initiated by the PSPP diphosphate returning back to the Mg2+ cluster for ionization, with the resultant DHS so formed being trapped in a surface pocket. This mechanism is supported by the observation that cationic inhibitors (of interest as antiinfectives) bind with their positive charge located in the same region as the cyclopropyl carbinyl group; that S-thiolo-diphosphates only inhibit when in the allylic site; activity results on 11 mutants show that both DXXXD conserved domains are essential for PSPP ionization; and the observation that head-to-tail isoprenoid synthases as well as terpene cyclases have ionization and alkene-donor sites which spatially overlap those found in CrtM.

AB - "Head-to-head" terpene synthases catalyze the first committed steps in sterol and carotenoid biosynthesis: the condensation of two isoprenoid diphosphates to form cyclopropylcarbinyl diphosphates, followed by ring opening. Here, we report the structures of Staphylococcus aureus dehydrosqualene synthase (CrtM) complexed with its reaction intermediate, presqualene diphosphate (PSPP), the dehydrosqualene (DHS) product, as well as a series of inhibitors. The results indicate that, on initial diphosphate loss, the primary carbocation so formed bends down into the interior of the protein to react with C2,3 double bond in the prenyl acceptor to form PSPP, with the lower two-thirds of both PSPP chains occupying essentially the same positions as found in the two farnesyl chains in the substrates. The second-half reaction is then initiated by the PSPP diphosphate returning back to the Mg2+ cluster for ionization, with the resultant DHS so formed being trapped in a surface pocket. This mechanism is supported by the observation that cationic inhibitors (of interest as antiinfectives) bind with their positive charge located in the same region as the cyclopropyl carbinyl group; that S-thiolo-diphosphates only inhibit when in the allylic site; activity results on 11 mutants show that both DXXXD conserved domains are essential for PSPP ionization; and the observation that head-to-tail isoprenoid synthases as well as terpene cyclases have ionization and alkene-donor sites which spatially overlap those found in CrtM.

KW - Drug discovery

KW - Quinuclidine

KW - Staphyloxanthin

KW - Triterpene

KW - X-ray crystallography

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JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

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