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

Research output: Contribution to journalArticle

42 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)21337-21342
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number50
DOIs
Publication statusPublished - 2010 Dec 14

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Diphosphates
Terpenes
Alkenes
Sterols
Carotenoids
Staphylococcus aureus
presqualene pyrophosphate
dehydrosqualene
Proteins

All Science Journal Classification (ASJC) codes

  • General

Cite this

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. In: Proceedings of the National Academy of Sciences of the United States of America. 2010 ; Vol. 107, No. 50. pp. 21337-21342.
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Lin, FY, Liu, CI, Liu, YL, Zhang, Y, Wang, K, Jeng, W-Y, 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, vol. 107, no. 50, pp. 21337-21342. https://doi.org/10.1073/pnas.1010907107

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.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, No. 50, 14.12.2010, p. 21337-21342.

Research output: Contribution to journalArticle

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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.

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