Localization and in-vivo characterization of Thapsia garganica CYP76AE2 indicates a role in thapsigargin biosynthesis

Trine Bundgaard Andersen; Karen Agatha Martinez-Swatson; Silas Anselm Rasmussen; Berin Alain Boughton; Kirsten Jørgensen; Johan Andersen-Ranberg; Nils Nyberg; Søren Brøgger Christensen; Henrik Toft Simonsen

doi:10.1104/pp.16.00055

Localization and in-vivo characterization of Thapsia garganica CYP76AE2 indicates a role in thapsigargin biosynthesis

Trine Bundgaard Andersen, Karen Agatha Martinez-Swatson, Silas Anselm Rasmussen, Berin Alain Boughton, Kirsten Jørgensen, Johan Andersen-Ranberg, Nils Nyberg, Søren Brøgger Christensen, Henrik Toft Simonsen^*

^*Corresponding author for this work

25 Citations (Scopus)

Abstract

The Mediterranean plant Thapsia garganica (dicot, Apiaceae), also known as deadly carrot, produces the highly toxic compound thapsigargin. This compound is a potent inhibitor of the sarcoplasmic-endoplasmic reticulum Ca²⁺ -ATPase calcium pump in mammals and is of industrial importance as the active moiety of the anticancer drug mipsagargin, currently in clinical trials. Knowledge of thapsigargin in planta storage and biosynthesis has been limited. Here, we present the putative second step in thapsigargin biosynthesis, by showing that the cytochrome P450 TgCYP76AE2, transiently expressed in Nicotiana benthamiana, converts epikunzeaol into epidihydrocostunolide. Furthermore, we show that thapsigargin is likely to be stored in secretory ducts in the roots. Transcripts from TgTPS2 (epikunzeaol synthase) and TgCYP76AE2 in roots were found only in the epithelial cells lining these secretory ducts. This emphasizes the involvement of these cells in the biosynthesis of thapsigargin. This study paves the way for further studies of thapsigargin biosynthesis.

Original language	English
Journal	Plant Physiology
Volume	174
Issue number	1
Pages (from-to)	56-72
Number of pages	17
ISSN	0032-0889
DOIs	https://doi.org/10.1104/pp.16.00055
Publication status	Published - 2017

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@article{7d93cee5e3f84fbfa0bc462584089ef1,

title = "Localization and in-vivo characterization of Thapsia garganica CYP76AE2 indicates a role in thapsigargin biosynthesis",

abstract = "The Mediterranean plant Thapsia garganica (dicot, Apiaceae), also known as deadly carrot, produces the highly toxic compound thapsigargin. This compound is a potent inhibitor of the sarcoplasmic-endoplasmic reticulum Ca2+ -ATPase calcium pump in mammals and is of industrial importance as the active moiety of the anticancer drug mipsagargin, currently in clinical trials. Knowledge of thapsigargin in planta storage and biosynthesis has been limited. Here, we present the putative second step in thapsigargin biosynthesis, by showing that the cytochrome P450 TgCYP76AE2, transiently expressed in Nicotiana benthamiana, converts epikunzeaol into epidihydrocostunolide. Furthermore, we show that thapsigargin is likely to be stored in secretory ducts in the roots. Transcripts from TgTPS2 (epikunzeaol synthase) and TgCYP76AE2 in roots were found only in the epithelial cells lining these secretory ducts. This emphasizes the involvement of these cells in the biosynthesis of thapsigargin. This study paves the way for further studies of thapsigargin biosynthesis.",

author = "Andersen, {Trine Bundgaard} and Martinez-Swatson, {Karen Agatha} and Rasmussen, {Silas Anselm} and Boughton, {Berin Alain} and Kirsten J{\o}rgensen and Johan Andersen-Ranberg and Nils Nyberg and Christensen, {S{\o}ren Br{\o}gger} and Simonsen, {Henrik Toft}",

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doi = "10.1104/pp.16.00055",

language = "English",

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T1 - Localization and in-vivo characterization of Thapsia garganica CYP76AE2 indicates a role in thapsigargin biosynthesis

AU - Andersen, Trine Bundgaard

AU - Martinez-Swatson, Karen Agatha

AU - Rasmussen, Silas Anselm

AU - Boughton, Berin Alain

AU - Jørgensen, Kirsten

AU - Andersen-Ranberg, Johan

AU - Nyberg, Nils

AU - Christensen, Søren Brøgger

AU - Simonsen, Henrik Toft

PY - 2017

Y1 - 2017

N2 - The Mediterranean plant Thapsia garganica (dicot, Apiaceae), also known as deadly carrot, produces the highly toxic compound thapsigargin. This compound is a potent inhibitor of the sarcoplasmic-endoplasmic reticulum Ca2+ -ATPase calcium pump in mammals and is of industrial importance as the active moiety of the anticancer drug mipsagargin, currently in clinical trials. Knowledge of thapsigargin in planta storage and biosynthesis has been limited. Here, we present the putative second step in thapsigargin biosynthesis, by showing that the cytochrome P450 TgCYP76AE2, transiently expressed in Nicotiana benthamiana, converts epikunzeaol into epidihydrocostunolide. Furthermore, we show that thapsigargin is likely to be stored in secretory ducts in the roots. Transcripts from TgTPS2 (epikunzeaol synthase) and TgCYP76AE2 in roots were found only in the epithelial cells lining these secretory ducts. This emphasizes the involvement of these cells in the biosynthesis of thapsigargin. This study paves the way for further studies of thapsigargin biosynthesis.

AB - The Mediterranean plant Thapsia garganica (dicot, Apiaceae), also known as deadly carrot, produces the highly toxic compound thapsigargin. This compound is a potent inhibitor of the sarcoplasmic-endoplasmic reticulum Ca2+ -ATPase calcium pump in mammals and is of industrial importance as the active moiety of the anticancer drug mipsagargin, currently in clinical trials. Knowledge of thapsigargin in planta storage and biosynthesis has been limited. Here, we present the putative second step in thapsigargin biosynthesis, by showing that the cytochrome P450 TgCYP76AE2, transiently expressed in Nicotiana benthamiana, converts epikunzeaol into epidihydrocostunolide. Furthermore, we show that thapsigargin is likely to be stored in secretory ducts in the roots. Transcripts from TgTPS2 (epikunzeaol synthase) and TgCYP76AE2 in roots were found only in the epithelial cells lining these secretory ducts. This emphasizes the involvement of these cells in the biosynthesis of thapsigargin. This study paves the way for further studies of thapsigargin biosynthesis.

U2 - 10.1104/pp.16.00055

DO - 10.1104/pp.16.00055

M3 - Journal article

C2 - 28275147

AN - SCOPUS:85018958858

SN - 0032-0889

VL - 174

SP - 56

EP - 72

JO - Plant Physiology

JF - Plant Physiology

IS - 1

ER -

Localization and in-vivo characterization of Thapsia garganica CYP76AE2 indicates a role in thapsigargin biosynthesis

Abstract

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