Analysis of 12-Oxo-phytodienoic Acid Enantiomers in Biological Samples by Capillary Gas Chromatography–Mass Spectrometry Using Cyclodextrin Stationary Phases

Laudert, Dietmar; Hennig, Peter; Stelmach, Boguslawa A.; Müller, Axel; Andert, Leo; Weiler, Elmar W.

doi:10.1006/abio.1997.2012

Cited by 55 publications

(48 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This suggests that multiple LOXs provide substrates for DA biosynthesis, similar to the complexity observed for JA biosynthesis (32). In contrast to the AOC-mediated enzymatic cyclization of 12,13-epoxyoctadecatrienoic acid yielding pure (9S,13S)-12-OPDA [i.e., cis(+)-12-OPDA] in potato and tomato, cis-10-OPEA and cis-10-OPDA are formed as minor racemic (9S,13S and 9R,13R) cyclization products of AOS-derived 9-allene oxides, which are predominantly hydrolyzed to α-ketols (29,30,33). Analysis of these relationships in maize unexpectedly revealed that inducible levels of cis-10-OPEA exceeded those of the predicted dominant α-ketol (9-hydroxy-10-oxo-12(Z)-octadecenoic acid) product by more than 15-fold (SI Appendix, Fig.…”

Section: Resultsmentioning

confidence: 99%

Maize death acids, 9-lipoxygenase–derived cyclopente(a)nones, display activity as cytotoxic phytoalexins and transcriptional mediators

Christensen

Huffaker

Kaplan

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

144

125

View full text Add to dashboard Cite

Plant damage promotes the interaction of lipoxygenases (LOXs) with fatty acids yielding 9-hydroperoxides, 13-hydroperoxides, and complex arrays of oxylipins. The action of 13-LOX on linolenic acid enables production of 12-oxo-phytodienoic acid (12-OPDA) and its downstream products, termed “jasmonates.” As signals, jasmonates have related yet distinct roles in the regulation of plant resistance against insect and pathogen attack. A similar pathway involving 9-LOX activity on linolenic and linoleic acid leads to the 12-OPDA positional isomer, 10-oxo-11-phytodienoic acid (10-OPDA) and 10-oxo-11-phytoenoic acid (10-OPEA), respectively; however, physiological roles for 9-LOX cyclopentenones have remained unclear. In developing maize (Zea mays) leaves, southern leaf blight (Cochliobolus heterostrophus) infection results in dying necrotic tissue and the localized accumulation of 10-OPEA, 10-OPDA, and a series of related 14- and 12-carbon metabolites, collectively termed “death acids.” 10-OPEA accumulation becomes wound inducible within fungal-infected tissues and at physiologically relevant concentrations acts as a phytoalexin by suppressing the growth of fungi and herbivores including Aspergillus flavus, Fusarium verticillioides, and Helicoverpa zea. Unlike previously established maize phytoalexins, 10-OPEA and 10-OPDA display significant phytotoxicity. Both 12-OPDA and 10-OPEA promote the transcription of defense genes encoding glutathione S transferases, cytochrome P450s, and pathogenesis-related proteins. In contrast, 10-OPEA only weakly promotes the accumulation of multiple protease inhibitor transcripts. Consistent with a role in dying tissue, 10-OPEA application promotes cysteine protease activation and cell death, which is inhibited by overexpression of the cysteine protease inhibitor maize cystatin-9. Unlike jasmonates, functions for 10-OPEA and associated death acids are consistent with specialized roles in local defense reactions.

show abstract

Section: Resultsmentioning

confidence: 99%

Maize death acids, 9-lipoxygenase–derived cyclopente(a)nones, display activity as cytotoxic phytoalexins and transcriptional mediators

Christensen

Huffaker

Kaplan

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

144

125

View full text Add to dashboard Cite

show abstract

“…Separations of the (þ)-and the (ÿ)-enantiomer of OPDA were made on a b-Dex120 column (30 m 3 0.25 mm 3 0.15 mm stationary-phase thickness) coated with 20% permethylb-cyclodextrin in SPB-35 (Supelco) with He carrier gas at 1 mL/min (gas prepressure of 80 kPa) using the following temperature program: injector temperature of 2208C, 1 min isothermally at 508C, with 108C/min up to 1908C, 75 min isothermally at 1908C, with 38C/min up to 2208C, 15 min isothermally at 2208C, and transfer line temperature 2208C. For details, see Laudert et al (1997) and Schaller et al (1998). Quantitation of AOC enzymatic activity was based on (þ)-OPDA-to-(ÿ)-OPDA ratios derived from integration of total ion current traces.…”

Section: Determination Of Total Enzymatic Activity Via Chiral Capillamentioning

confidence: 99%

“…The extremely short halflife of allene oxides (half-time <30 s in water; Brash et al, 1988) and the optical purity of natural OPDA suggest a tight coupling of the AOS and AOC reactions, possibly in a synthase-cyclase complex (Stenzel et al, 2003b). Coupling of the two reactions is also observed in vitro: AOC from potato (Solanum tuberosum) or recombinant Arabidopsis thaliana AOC2 in combination with recombinant Arabidopsis AOS resulted in the production of highly asymmetrical cis-OPDA consisting nearly exclusively of the (9S,13S)-enantiomer (Laudert et al, 1997). Recently, we demonstrated that a physical interaction of AOS and AOC is not essential for the cyclization reaction, but the yield of OPDA formation and optical purity of the product is diminished when separating both enzymes from each other (P. Zerbe, E.W.…”

Section: Introductionmentioning

confidence: 96%

“…Reduction of the 10,11-double bond by an NADPH-dependent OPDA reductase then yields 3-oxo-2(29(Z)-pentenyl)-cyclopentane-1-octanoic acid , which is believed to undergo three cycles of b-oxidation to yield the end product of the pathway [i.e., JA with (3R,7S)-configuration; (þ)-7-iso-JA] (Vick and Zimmerman, 1984). The biosynthesis of JA appears to involve two different compartments: the conversion of LA to OPDA is localized in the chloroplasts (Vick and Zimmerman, 1987;Song et al, 1993;Laudert et al, 1997), while the reduction of OPDA to OPC-8:0 Weiler, 1997a, 1997b;Schaller et al, 2000;Strassner et al, 2002) and the three steps of b-oxidation (i.e., conversion of OPC-8:0 to JA) occur in peroxisomes (Gerhardt, 1983;Vick and Zimmerman, 1984;Li et al, 2005). Like JA, OPDA may accumulate to substantial amounts in plant tissue .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The Crystal Structure of Arabidopsis thaliana Allene Oxide Cyclase: Insights into the Oxylipin Cyclization Reaction

2006

View full text Add to dashboard Cite

We describe the crystallization and structure elucidation of Arabidopsis thaliana allene oxide cyclase 2 (AOC2), a key enzyme in the biosynthesis of jasmonates. In a coupled reaction with allene oxide synthase, AOC2 releases the first cyclic and biologically active metabolite, 12-oxo-phytodienoic acid (OPDA). AOC2 (AT3G25770) folds into an eight-stranded antiparallel b-barrel with a C-terminal partial helical extension. The protein forms a hydrophobic binding cavity with two distinct polar patches. AOC2 is trimeric in crystals, in vitro and in planta. Based on the observed folding pattern, we assigned AOC2 as a low molecular weight member of the lipocalin family with enzymatic activity in plants. We determined the binding position of the competitive inhibitor vernolic acid (a substrate analog) in the binding pocket. Based on models for bound substrate 12,13-epoxy-9,11,15-octadecatrienoic acid and product OPDA, we propose a reaction scheme that explains the influence of the C15 double bond on reactivity. Reaction is promoted by anchimeric assistance through a conserved Glu residue. The transition state with a pentadienyl carbocation and an oxyanion is stabilized by a strongly bound water molecule and favorable p-p interactions with aromatic residues in the cavity. Stereoselectivity results from steric restrictions to the necessary substrate isomerizations imposed by the protein.

show abstract

“…1), and a cDNA encoding an OPR was first cloned from Arabidopsis, using sequence of the enzyme purified from Corydalis sempervirens (16). However, subsequent studies (17) showed that the Arabidopsis gene, OPR1, encodes a protein with almost no activity against 9S,13S-OPDA, which is believed to be the principal isomer in plants (18). At the same time, a second, separable OPR activity from C. sempervirens was shown to have the characteristics expected of an isozyme involved in JA synthesis in that it efficiently converted 9S,13S-OPDA to 3-oxo-2(2Ј[Z]-pentenyl)-cyclopentane-1-octanoic acid (OPC:8) (17), which is hypothesized to be converted to 3R,7S-JA [(ϩ)-7-iso-JA] by three cycles of ␤-oxidation (19).…”

mentioning

confidence: 99%

The Arabidopsis male-sterile mutant, opr3 , lacks the 12-oxophytodienoic acid reductase required for jasmonate synthesis

Stintzi

Browse

2000

Proc. Natl. Acad. Sci. U.S.A.

741

733

View full text Add to dashboard Cite

Jasmonic acid (JA) and its precursor 12-oxophytodienoic acid (OPDA) act as plant growth regulators and mediate responses to environmental cues. To investigate the role of these oxylipins in anther and pollen development, we characterized a T-DNA-tagged, male-sterile mutant of Arabidopsis, opr3. The opr3 mutant plants are sterile but can be rendered fertile by exogenous JA but not by OPDA. Cloning of the mutant locus indicates that it encodes an isozyme of 12-oxophytodienoate reductase, designated OPR3. All of the defects in opr3 are alleviated by transformation of the mutant with an OPR3 cDNA. Our results indicate that JA and not OPDA is the signaling molecule that induces and coordinates the elongation of the anther filament, the opening of the stomium at anthesis, and the production of viable pollen. Just as importantly, our data demonstrate that OPR3 is the only isoform of OPR capable of reducing the correct stereoisomer of OPDA to produce JA required for male gametophyte development.

show abstract

Analysis of 12-Oxo-phytodienoic Acid Enantiomers in Biological Samples by Capillary Gas Chromatography–Mass Spectrometry Using Cyclodextrin Stationary Phases

Cited by 55 publications

References 41 publications

Maize death acids, 9-lipoxygenase–derived cyclopente(a)nones, display activity as cytotoxic phytoalexins and transcriptional mediators

Maize death acids, 9-lipoxygenase–derived cyclopente(a)nones, display activity as cytotoxic phytoalexins and transcriptional mediators

The Crystal Structure of Arabidopsis thaliana Allene Oxide Cyclase: Insights into the Oxylipin Cyclization Reaction

The Arabidopsis male-sterile mutant, opr3 , lacks the 12-oxophytodienoic acid reductase required for jasmonate synthesis

Contact Info

Product

Resources

About