X‐ray structure of <i>Arabidopsis</i> At2g06050, 12‐oxophytodienoate reductase isoform 3

Malone, Thomas E.; Madson, Stacey E.; Wrobel, Russell L.; Jeon, Won Bae; Rosenberg, Nathan; Johnson, Kenneth A.; Bingman, C.A.; Smith, David W.; Phillips, George N.; Markley, John L.; Fox, Brian G.

doi:10.1002/prot.20162

Cited by 21 publications

(22 citation statements)

References 15 publications

(14 reference statements)

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“…3b). This structural correspondence is also found for nearly all structurally characterized members of the OYE family (data not shown) with the exception of His-188, which is replaced by asparagine in several enzymes (19,20,(25)(26)(27)(28). The corresponding atoms Asn N␦2 and His N␦1 that bind the carbonyl oxygen overlay perfectly, however, indicating a similar effect on substrate activation and positioning.…”

Section: Resultssupporting

confidence: 66%

“…The structural and biochemical data presented in this study show that LeOPR3 from tomato can exist as a self-inhibited dimer. In this context, it seems surprising that the recently reported structure of AtOPR3 does not show an obvious dimer (28). Yet, the two ''monomers'' observed in the asymmetric unit of AtOPR3 crystals can be superimposed with the corresponding protomers of the LeOPR3 dimer, suggesting a similar dimeric structure for AtOPR3.…”

Section: Homodimerization In Solutionmentioning

confidence: 86%

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Crystal structure of 12-oxophytodienoate reductase 3 from tomato: Self-inhibition by dimerization

Breithaupt

Kurzbauer

Lilie

et al. 2006

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

118

109

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12-Oxophytodienoate reductase (OPR) 3, a homologue of old yellow enzyme (OYE), catalyzes the reduction of 9S,13S-12-oxophytodienoate to the corresponding cyclopentanone, which is subsequently converted to the plant hormone jasmonic acid (JA). JA and JA derivatives, as well as 12-oxophytodienoate and related cyclopentenones, are known to regulate gene expression in plant development and defense. Together with other oxygenated fatty acid derivatives, they form the oxylipin signature in plants, which resembles the pool of prostaglandins in animals. Here, we report the crystal structure of OPR3 from tomato and of two OPR3 mutants. Although the catalytic residues of OPR3 and related OYEs are highly conserved, several characteristic differences can be discerned in the substrate-binding regions, explaining the remarkable substrate stereoselectivity of OPR isozymes. Interestingly, OPR3 crystallized as an extraordinary self-inhibited dimer. Mutagenesis studies and biochemical analysis confirmed a weak dimerization of OPR3 in vitro, which correlated with a loss of enzymatic activity. Based on structural data of OPR3, a putative mechanism for a strong and reversible dimerization of OPR3 in vivo that involves phosphorylation of OPR3 is suggested. This mechanism could contribute to the shaping of the oxylipin signature, which is critical for fine-tuning gene expression in plants.flavoprotein ͉ jasmonic acid biosynthesis ͉ plant defense ͉ oxylipin signature ͉ 12-oxophytodienoic acid F lavoproteins catalyze a wide variety of essential biochemical reactions, including electron transfer, dehydrogenation, and hydroxylation reactions. Old yellow enzyme (OYE), the first flavin-dependent enzyme identified (1), and homologues of OYE from bacteria, yeast, and plants are able to reduce the CAC double bond of ␣,␤-unsaturated carbonyl compounds, an activity that is rather uncommon for flavoenzymes (2, 3). This reaction has been shown to proceed by a ping-pong bi-bi mechanism, during which the FMN cofactor is reduced by NAD(P)H before the substrate is bound and reduced by hydride transfer to the substrate's C ␤ (4). Despite extensive efforts, the physiological substrate has been revealed only for one member of the OYE family, the plant enzyme 12-oxophytodienoate reductase (OPR) 3, which catalyzes one step in the biosynthesis of the plant hormone jasmonic acid (JA) (5, 6). JA and JA derivatives act as signaling compounds in the defense response against herbivores and pathogens and are involved in the regulation of various developmental processes, such as fruit ripening, pollen maturation, and senescence (7,8). JA is synthesized from ␣-linolenic acid, which is oxidized and cyclized, resulting in the cyclopentenone 9S,13S-12-oxophytodienoate (9S,13S-OPDA). OPR3 reduces 9S,13S-OPDA to the corresponding cyclopentanone, which is converted to JA by repeated ␤-oxidation ( Fig. 1) (9). Several OPR isozymes have been identified in plants, including 3 isoforms in Lycopersicon esculentum, 5 in Arabidopsis thaliana, and 13 in Oryza sativa (10, 11). O...

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Section: Resultssupporting

confidence: 66%

Section: Homodimerization In Solutionmentioning

confidence: 86%

Crystal structure of 12-oxophytodienoate reductase 3 from tomato: Self-inhibition by dimerization

Breithaupt

Kurzbauer

Lilie

et al. 2006

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

118

109

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“…The overall structure of YqjM strongly resembles the structures of other OYE homologs (13)(14)(15)(16)(17). The closest structural relative was found to be pentaerythritol tetranitrate (PETN) reductase from Enterobacter cloacae where 278 residues could be aligned with a root mean square deviation (r.m.s.d.)…”

Section: Resultsmentioning

confidence: 86%

The 1.3 Å Crystal Structure of the Flavoprotein YqjM Reveals a Novel Class of Old Yellow Enzymes

Kitzing

Fitzpatrick

Wilken

et al. 2005

Journal of Biological Chemistry

130

198

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Here we report the crystal structure of YqjM, a homolog of Old Yellow Enzyme (OYE) that is involved in the oxidative stress response of Bacillus subtilis. In addition to the oxidized and reduced enzyme form, the structures of complexes with p-hydroxybenzaldehyde and pnitrophenol, respectively, were solved. As for other OYE family members, YqjM folds into a (␣/␤) 8 -barrel and has one molecule of flavin mononucleotide bound non-covalently at the COOH termini of the ␤-sheet. Most of the interactions that control the electronic properties of the flavin mononucleotide cofactor are conserved within the OYE family. However, in contrast to all members of the OYE family characterized to date, YqjM exhibits several unique structural features. For example, the enzyme exists as a homotetramer that is assembled as a dimer of catalytically dependent dimers. Moreover, the protein displays a shared active site architecture where an arginine finger (Arg 336 ) at the COOH terminus of one monomer extends into the active site of the adjacent monomer and is directly involved in substrate recognition. Another remarkable difference in the binding of the ligand in YqjM is represented by the contribution of the NH 2 -terminal Tyr 28 instead of a COOH-terminal tyrosine in OYE and its homologs. The structural information led to a specific data base search from which a new class of OYE oxidoreductases was identified that exhibits a strict conservation of active site residues, which are critical for this subfamily, most notably Cys 26 , Tyr 28 , Lys 109 , and Arg 336 . Therefore, YqjM is the first representative of a new bacterial subfamily of OYE homologs.

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“…S2). Although it is unlikely that reductases other than OPR3 convert OPDA to OPC:8 (Breithaupt et al, 2001(Breithaupt et al, , 2006Malone et al, 2005;Hall et al, 2007Hall et al, , 2008Beynon, et al, 2009;Schaller and Stintzi, 2009), whether OPR2 or other OPRs are involved in the production of JAs and thus play a role in fungal resistance remains to be determined.…”

Section: Opr3 Is Not a Loss-of-function Mutantmentioning

confidence: 99%

Intronic T-DNA Insertion Renders Arabidopsisopr3a Conditional Jasmonic Acid-Producing Mutant

Chehab

Savchenko

et al. 2011

Plant Physiology

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Jasmonic acid and its derived metabolites (JAs) orchestrate plant defense against insects and fungi. 12-Oxo-phytodienoic acid (OPDA), a JA precursor, has also been implicated in plant defense. We sought to define JAs and OPDA functions through comparative defense susceptibility characteristics of three Arabidopsis (Arabidopsis thaliana) genotypes: aos, lacking JAs and OPDA; opda reductase3 (opr3), deficient in JA production but can accumulate OPDA; and transgenics that overexpress OPR3. opr3, like aos, is susceptible to cabbage loopers (Trichoplusia ni) but, relative to aos, opr3 has enhanced resistance to a necrotrophic fungus. Gas chromatography-mass spectrometry reveals that opr3 produces OPDA but no detectable JAs following wounding and looper infestation; unexpectedly, substantial levels of JAs accumulate in opr3 upon fungal infection. Full-length OPR3 transcripts accumulate in fungal-infected opr3, potentially through splicing of the T-DNA containing intron. Fungal resistance correlates with levels of JAs not OPDA; therefore, opr3 resistance to some pests is likely due to JA accumulation, and signaling activities ascribed to OPDA should be reassessed because opr3 can produce JAs. Together these data (1) reinforce the primary role JAs play in plant defense against insects and necrotrophic fungi, (2) argue for a reassessment of signaling activities ascribed to OPDA, and (3) provide evidence that mutants with intron insertions can retain gene function.

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X‐ray structure of Arabidopsis At2g06050, 12‐oxophytodienoate reductase isoform 3

Cited by 21 publications

References 15 publications

Crystal structure of 12-oxophytodienoate reductase 3 from tomato: Self-inhibition by dimerization

Crystal structure of 12-oxophytodienoate reductase 3 from tomato: Self-inhibition by dimerization

The 1.3 Å Crystal Structure of the Flavoprotein YqjM Reveals a Novel Class of Old Yellow Enzymes

Intronic T-DNA Insertion Renders Arabidopsisopr3a Conditional Jasmonic Acid-Producing Mutant

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