AtGH3.10 and JAR1 Produce 12-Hydroxyjasmonoyl-l-isoleucine from 12-Hydroxyjasmonic Acid in Arabidopsis thaliana

Katsunari Oki; Akane Enoki; Yoshitaka Yokota; Taiki Kurihara; Takafumi Shimizu; Wataru Saburi; Takayuki Tohge; Haruhide Mori; Guido Van den Ackerveken; Naoki Kitaoka; Hideyuki Matsuura

doi:10.1002/cbic.202500151

AtGH3.10 and JAR1 Produce 12-Hydroxyjasmonoyl-l-isoleucine from 12-Hydroxyjasmonic Acid in Arabidopsis thaliana

Katsunari Oki, Akane Enoki, Yoshitaka Yokota, Taiki Kurihara, Takafumi Shimizu, Wataru Saburi, Takayuki Tohge, Haruhide Mori, Guido Van den Ackerveken, Naoki Kitaoka^*, Hideyuki Matsuura^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Jasmonates are plant hormones that regulate plant defense and development. 7-iso-Jasmonoyl-l-isoleucine (JA-Ile) is a representative active jasmonate which is biosynthesized from 7-iso-jasmonic acid (JA) by the jasmonoyl-amido synthases JASMONATE RESISTANT 1 (JAR1) and AtGH3.10 in Arabidopsis thaliana. 12-Hydroxy-7-iso-jasmonoyl-l-isoleucine (12-OH-JA-Ile) is another active jasmonate, and 12-hydroxylation of JA-Ile is considered the major biosynthetic pathway toward 12-OH-JA-Ile. Previous report elucidated that recombinant JAR1 showed a weak activity against 12-hydroxy-7-iso-jasmonic acid (12-OH-JA). However, the direct conversion from 12-OH-JA to 12-OH-JA-Ile in planta and the enzyme activity of AtG3.10 against 12-OH-JA have never been reported. Herein, a feeding experiment with deuterated 12-OH-JA confirms the direct conversion of 12-OH-JA to 12-OH-JA-Ile in wild-type Arabidopsis plants. The conversion from 12-OH-JA to 12-OH-JA-Ile is not observed in jar1 gh3.10 double mutant, suggesting that 12-OH-JA is converted to 12-OH-JA-Ile by JAR1 and AtGH3.10. Notably, enzyme assays show that the catalytic efficiency with 12-OH-JA for AtGH3.10 is higher than those with JA for AtGH3.10 and with 12-OH-JA for JAR1. Comparative analysis of JAR1 and AtGH3.10 structures and site-directed mutation analysis reveals that Ser120 in AtGH3.10 is the key amino acid residue responsible for its high catalytic efficiency against 12-OH-JA.

Original language	English
Article number	e202500151
Journal	ChemBioChem
Volume	26
Issue number	19
Early online date	11 Sept 2025
DOIs	https://doi.org/10.1002/cbic.202500151
Publication status	Published - 3 Oct 2025

Bibliographical note

Publisher Copyright:
© 2025 Wiley-VCH GmbH.

Funding

K.O., A.E., and Y.Y. contributed equally to this work. This work was supported by a grant from the Japan Society for the Promotion of Science KAKENHI (23K05047) to N.K. This study was supported in part by Grants‐in‐Aid for Regional R&D Proposal‐Based Program from Northern Advancement Center for Science & Technology of Hokkaido Japan. This work was also supported by the research grant from Japan Society for Bioscience, Biotechnology, and Agrobiochemistry. The authors thank Dr. Eri Fukushi and Mr. Yusuke Takata (GC‐MS &NMR Laboratory, Research Faculty of Agriculture, Hokkaido University) for their assistance in obtaining the spectroscopy data. The authors also thank Ms. Kanako Tannai (Research Faculty of Agriculture, Hokkaido University) and Ms. Asako Kawabe (Research Faculty of Agriculture, Hokkaido University) for their technical assistance. The authors thank ZEON Corporation for providing the methyl jasmonate. The UPLC MS/MS system used in this study is supported by funds from Program for Forming Japan's Peak Research Universities (J‐PEAKS, Japan Society for The Promotion of Science) and housed at the Research Faculty of Agriculture, Hokkaido University. The authors thank Editage [ http://www.editage.com ] for their English language service in editing and reviewing this manuscript.

Funders	Funder number
Peak Research Universities
Japan Society for Bioscience, Biotechnology, and Agrochemistry
Northern Advancement Center for Science and Technology
Japan Society for the Promotion of Science	23K05047

Keywords

biosynthesis
gretchen Hagen 3
jasmonates
jasmonoyl-amido synthase
phytochemistry

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@article{a7fe3059c489409eb51021477eba1fee,

title = "AtGH3.10 and JAR1 Produce 12-Hydroxyjasmonoyl-l-isoleucine from 12-Hydroxyjasmonic Acid in Arabidopsis thaliana",

abstract = "Jasmonates are plant hormones that regulate plant defense and development. 7-iso-Jasmonoyl-l-isoleucine (JA-Ile) is a representative active jasmonate which is biosynthesized from 7-iso-jasmonic acid (JA) by the jasmonoyl-amido synthases JASMONATE RESISTANT 1 (JAR1) and AtGH3.10 in Arabidopsis thaliana. 12-Hydroxy-7-iso-jasmonoyl-l-isoleucine (12-OH-JA-Ile) is another active jasmonate, and 12-hydroxylation of JA-Ile is considered the major biosynthetic pathway toward 12-OH-JA-Ile. Previous report elucidated that recombinant JAR1 showed a weak activity against 12-hydroxy-7-iso-jasmonic acid (12-OH-JA). However, the direct conversion from 12-OH-JA to 12-OH-JA-Ile in planta and the enzyme activity of AtG3.10 against 12-OH-JA have never been reported. Herein, a feeding experiment with deuterated 12-OH-JA confirms the direct conversion of 12-OH-JA to 12-OH-JA-Ile in wild-type Arabidopsis plants. The conversion from 12-OH-JA to 12-OH-JA-Ile is not observed in jar1 gh3.10 double mutant, suggesting that 12-OH-JA is converted to 12-OH-JA-Ile by JAR1 and AtGH3.10. Notably, enzyme assays show that the catalytic efficiency with 12-OH-JA for AtGH3.10 is higher than those with JA for AtGH3.10 and with 12-OH-JA for JAR1. Comparative analysis of JAR1 and AtGH3.10 structures and site-directed mutation analysis reveals that Ser120 in AtGH3.10 is the key amino acid residue responsible for its high catalytic efficiency against 12-OH-JA.",

keywords = "biosynthesis, gretchen Hagen 3, jasmonates, jasmonoyl-amido synthase, phytochemistry",

author = "Katsunari Oki and Akane Enoki and Yoshitaka Yokota and Taiki Kurihara and Takafumi Shimizu and Wataru Saburi and Takayuki Tohge and Haruhide Mori and \{Van den Ackerveken\}, Guido and Naoki Kitaoka and Hideyuki Matsuura",

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month = oct,

day = "3",

doi = "10.1002/cbic.202500151",

language = "English",

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T1 - AtGH3.10 and JAR1 Produce 12-Hydroxyjasmonoyl-l-isoleucine from 12-Hydroxyjasmonic Acid in Arabidopsis thaliana

AU - Oki, Katsunari

AU - Enoki, Akane

AU - Yokota, Yoshitaka

AU - Kurihara, Taiki

AU - Shimizu, Takafumi

AU - Saburi, Wataru

AU - Tohge, Takayuki

AU - Mori, Haruhide

AU - Van den Ackerveken, Guido

AU - Kitaoka, Naoki

AU - Matsuura, Hideyuki

PY - 2025/10/3

Y1 - 2025/10/3

N2 - Jasmonates are plant hormones that regulate plant defense and development. 7-iso-Jasmonoyl-l-isoleucine (JA-Ile) is a representative active jasmonate which is biosynthesized from 7-iso-jasmonic acid (JA) by the jasmonoyl-amido synthases JASMONATE RESISTANT 1 (JAR1) and AtGH3.10 in Arabidopsis thaliana. 12-Hydroxy-7-iso-jasmonoyl-l-isoleucine (12-OH-JA-Ile) is another active jasmonate, and 12-hydroxylation of JA-Ile is considered the major biosynthetic pathway toward 12-OH-JA-Ile. Previous report elucidated that recombinant JAR1 showed a weak activity against 12-hydroxy-7-iso-jasmonic acid (12-OH-JA). However, the direct conversion from 12-OH-JA to 12-OH-JA-Ile in planta and the enzyme activity of AtG3.10 against 12-OH-JA have never been reported. Herein, a feeding experiment with deuterated 12-OH-JA confirms the direct conversion of 12-OH-JA to 12-OH-JA-Ile in wild-type Arabidopsis plants. The conversion from 12-OH-JA to 12-OH-JA-Ile is not observed in jar1 gh3.10 double mutant, suggesting that 12-OH-JA is converted to 12-OH-JA-Ile by JAR1 and AtGH3.10. Notably, enzyme assays show that the catalytic efficiency with 12-OH-JA for AtGH3.10 is higher than those with JA for AtGH3.10 and with 12-OH-JA for JAR1. Comparative analysis of JAR1 and AtGH3.10 structures and site-directed mutation analysis reveals that Ser120 in AtGH3.10 is the key amino acid residue responsible for its high catalytic efficiency against 12-OH-JA.

AB - Jasmonates are plant hormones that regulate plant defense and development. 7-iso-Jasmonoyl-l-isoleucine (JA-Ile) is a representative active jasmonate which is biosynthesized from 7-iso-jasmonic acid (JA) by the jasmonoyl-amido synthases JASMONATE RESISTANT 1 (JAR1) and AtGH3.10 in Arabidopsis thaliana. 12-Hydroxy-7-iso-jasmonoyl-l-isoleucine (12-OH-JA-Ile) is another active jasmonate, and 12-hydroxylation of JA-Ile is considered the major biosynthetic pathway toward 12-OH-JA-Ile. Previous report elucidated that recombinant JAR1 showed a weak activity against 12-hydroxy-7-iso-jasmonic acid (12-OH-JA). However, the direct conversion from 12-OH-JA to 12-OH-JA-Ile in planta and the enzyme activity of AtG3.10 against 12-OH-JA have never been reported. Herein, a feeding experiment with deuterated 12-OH-JA confirms the direct conversion of 12-OH-JA to 12-OH-JA-Ile in wild-type Arabidopsis plants. The conversion from 12-OH-JA to 12-OH-JA-Ile is not observed in jar1 gh3.10 double mutant, suggesting that 12-OH-JA is converted to 12-OH-JA-Ile by JAR1 and AtGH3.10. Notably, enzyme assays show that the catalytic efficiency with 12-OH-JA for AtGH3.10 is higher than those with JA for AtGH3.10 and with 12-OH-JA for JAR1. Comparative analysis of JAR1 and AtGH3.10 structures and site-directed mutation analysis reveals that Ser120 in AtGH3.10 is the key amino acid residue responsible for its high catalytic efficiency against 12-OH-JA.

KW - biosynthesis

KW - gretchen Hagen 3

KW - jasmonates

KW - jasmonoyl-amido synthase

KW - phytochemistry

UR - https://www.scopus.com/pages/publications/105015529543

U2 - 10.1002/cbic.202500151

DO - 10.1002/cbic.202500151

M3 - Article

C2 - 40931695

SN - 1439-4227

VL - 26

JO - ChemBioChem

JF - ChemBioChem

IS - 19

M1 - e202500151

ER -

AtGH3.10 and JAR1 Produce 12-Hydroxyjasmonoyl-l-isoleucine from 12-Hydroxyjasmonic Acid in Arabidopsis thaliana

Abstract

Bibliographical note

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Keywords

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