Structural insights into the evolutionary paths of oxylipin biosynthetic enzymes

ABSTRACT The oxylipin pathway generates not only prostaglandin-like jasmonates but also green leaf volatiles (GLVs), which confer characteristic aromas to fruits and vegetables. Although

allene oxide synthase (AOS) and hydroperoxide lyase are atypical cytochrome P450 family members involved in the synthesis of jasmonates and GLVs, respectively, it is unknown how these

enzymes rearrange their hydroperoxide substrates into different products. Here we present the crystal structures of _Arabidopsis thaliana_ AOS, free and in complex with substrate or

intermediate analogues. The structures reveal an unusual active site poised to control the reactivity of an epoxyallylic radical and its cation by means of interactions with an aromatic

π-system. Replacing the amino acid involved in these steps by a non-polar residue markedly reduces AOS activity and, unexpectedly, is both necessary and sufficient for converting AOS into a

GLV biosynthetic enzyme. Furthermore, by combining our structural data with bioinformatic and biochemical analyses, we have discovered previously unknown hydroperoxide lyase in plant

growth-promoting rhizobacteria, AOS in coral, and epoxyalcohol synthase in amphioxus. These results indicate that oxylipin biosynthetic genes were present in the last common ancestor of

plants and animals, but were subsequently lost in all metazoan lineages except Placozoa, Cnidaria and Cephalochordata. Access through your institution Buy or subscribe This is a preview of

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ADDITIONAL ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS COMPLETE BIOSYNTHETIC PATHWAY

OF FUROCHROMONES IN _SAPOSHNIKOVIA DIVARICATA_ AND ITS EVOLUTIONARY MECHANISM IN APIACEAE PLANTS Article Open access 01 April 2025 THE EVOLUTIONARY ORIGIN OF NATURALLY OCCURRING

INTERMOLECULAR DIELS-ALDERASES FROM _MORUS ALBA_ Article Open access 20 March 2024 THE GRADUAL ESTABLISHMENT OF COMPLEX COUMARIN BIOSYNTHETIC PATHWAY IN APIACEAE Article Open access 10

August 2024 ACCESSION CODES PRIMARY ACCESSIONS GENBANK/EMBL/DDBJ * EU541487 * EU555186 * EU887514 PROTEIN DATA BANK * 2RCH * 2RCL * 2RCM * 3CLI * 3DSI * 3DSJ * 3DSK DATA DEPOSITS Coordinates

and structure factors have been deposited in the RCSB Protein Data Bank under the following accession codes: 3CLI (At-AOS), 3DSI (At-AOS in complex with 13-HOT), 2RCH (At-AOS in complex

with 13-HOD), 2RCL (At-AOS in complex with 12_R_,13_S_-vernolic acid), 2RCM (At-AOS(F137L)), 3DSJ (At-AOS(F137L) in complex with 13-HOD), and 3DSK (At-AOS(F137L) in complex with

12_R_,13_S_-vernolic acid). Nucleotide sequences of Ap-AOS, Bf-EAS and Mn-HPL have been deposited in Genbank under accession numbers EU541487, EU555186 and EU887514, respectively. REFERENCES

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<http://www.pymol.org> (DeLano Scientific, 2002) Download references ACKNOWLEDGEMENTS C.S.R. is grateful to the late N. Natarajaratnam and the late T. Traylor for

encouragement; R. Cudney for nonanoyl-_N_-hydroxyethylglucamide (HEGA-9); K. Matsui for tomato _HPL_ complementary DNA; K. Back and D. Park for _Arabidopsis_ and rice cDNA libraries; L.

Holland and J. Langeland for the amphioxus cDNA library; M. Medina for _A. palmata_ expressed sequence tags; C. Marx and L. Moulin for _M. nodulans_ cells; R. Müller for _S. aurantiaca_

genomic DNA; L. Roman and B. Masters for the pCWori+ vector; J. Navarro for the single crystal microspectrophotometer; T. Doukov, S. Soltis, A. Cohen and J. Charles for help with acquiring

electronic absorption spectra of single crystals; S. Veeraraghavan for generating Fig. 5; R. Bach, T. Bach, A. Beckwith, W. Bernhard, D. Curran, A. Davies, T. Dibble, J. Finnerty, D.

Fleischman, J. Froehlich, J. Groves, L. Holland, P. Holland, J. Howieson, H. Kaplan, D. Nelson, M. Newcomb, P. Ortiz de Montellano, N. Porter, T. Poulos, M. Sibi, S. Veeraraghavan and D.

Whalen for discussions; Joint Genome Institute for access to sequence data; Stanford Synchrotron Radiation Laboratories (beam lines 9–2 and 11–1, T. Doukov and L. Dunn) and the Advanced

Light Source (beam line 8.3.1, J. Holton, G. Meigs and J. Tanamachi; beam lines 8.2.1 and 8.2.2, C. Ralston) for beam time and assistance. This work is supported by Pew Charitable Trusts

through a Pew Scholar Award (C.S.R.), The Robert A. Welch Foundation (C.S.R.), The National Institutes of Health (C.S.R.), a Beginning Grant in Aid from the American Heart Association

(D.-S.L.), and an INSERM Avenir Grant sponsored by La Fondation pour la Recherche Médicale (P.N.). AUTHOR CONTRIBUTIONS C.S.R. designed the research. D.-S.L. overexpressed and purified all

the proteins used in this work; D.-S.L. and P.N. measured enzyme kinetic data; D.-S.L. generated the crystals; D.-S.L. and C.S.R. collected X-ray diffraction data; P.N. and C.S.R. solved the

structures; P.N. did structure refinements; C.S.R. carried out bioinformatic and phylogenetic analyses; M.H. performed GC–MS and radio-HPLC measurements, and determined the structures of

the reaction products; and C.S.R. wrote the paper. All authors discussed the results and commented on the manuscript. AUTHOR INFORMATION Author notes * Dong-Sun Lee and Pierre Nioche: These

authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston, Texas 77030, USA, Dong-Sun

Lee & C. S. Raman * Université Paris Descartes, INSERM UMR-S 747, 75270 Paris cedex 06, France , Pierre Nioche * Division of Physiological Chemistry II, Department of Medical

Biochemistry & Biophysics, Karolinska Institutet, S-17177 Stockholm, Sweden Mats Hamberg Authors * Dong-Sun Lee View author publications You can also search for this author inPubMed 

Google Scholar * Pierre Nioche View author publications You can also search for this author inPubMed Google Scholar * Mats Hamberg View author publications You can also search for this

author inPubMed Google Scholar * C. S. Raman View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to C. S. Raman.

SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION The file contains Supplementary Figures 1- 34 with Legends, Supplementary Tables 1-6, Supplementary Discussion and additional references.

(PDF 7022 kb) SUPPLEMENTARY MOVIE 1 The file contains Supplementary Movie 1. This movie reveals how the substrate is recognized at the active site of Arabidopsis thaliana allene oxide

synthase (At-AOS) (SWF 9060 kb) SUPPLEMENTARY MOVIE 2 The file contains Supplementary Movie 2. This movie shows the mode of interaction between the reaction intermediate analog and At-AOS.

(SWF 8652 kb) POWERPOINT SLIDES POWERPOINT SLIDE FOR FIG. 1 POWERPOINT SLIDE FOR FIG. 2 POWERPOINT SLIDE FOR FIG. 3 POWERPOINT SLIDE FOR FIG. 4 POWERPOINT SLIDE FOR FIG. 5 POWERPOINT SLIDE

FOR FIG. 6 RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Lee, DS., Nioche, P., Hamberg, M. _et al._ Structural insights into the evolutionary paths of

oxylipin biosynthetic enzymes. _Nature_ 455, 363–368 (2008). https://doi.org/10.1038/nature07307 Download citation * Received: 14 November 2007 * Accepted: 01 August 2008 * Published: 20

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