Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Aug 8;15(1):6714.
doi: 10.1038/s41467-024-50705-2.

Reply to: LsBOS utilizes oxalyl-CoA produced by LsAAE3 to synthesize β-ODAP in grass pea

Affiliations

Reply to: LsBOS utilizes oxalyl-CoA produced by LsAAE3 to synthesize β-ODAP in grass pea

Anne Edwards et al. Nat Commun. .
No abstract available

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1. LsAAE3 enzyme assays of α/β-L-ODAP formation.
Shown are measurements of the supernatant after removal of LsAAE3, removal of LsAAE3 followed by addition of LsBOS and the coupled reaction in the presence of both LsAAE3 and LsBOS. a concentrations of α- and β-ODAP produced. b Western blot showing LsAAE3 in the initial reaction (lane 1) and absent in the supernatant following filtration and addition of LsBOS (lanes 2–4), alongside Biorad Precision Plus Marker. c LC-MS profiles of assays showing no detectable oxalyl-CoA. The bottom trace shows oxalyl-CoA produced in vitro using purified LsAAE3 as described in Edwards et al.. Source data are provided as a Source Data file.
Fig. 2
Fig. 2. α/β-L-ODAP formation in the absence of LsBOS.
LCMS spectra show α- and β-ODAP formation in vitro using oxalate and L-DAP in combination with LsAAE3 and 20 nM desulpho-CoA (track 1); LsAAE3 and 20 nM CoA (track 2); LsAAE3 only (track 3); or α/β-L-ODAP standard. Source data are provided as a Source Data file.
Fig. 3
Fig. 3. Detection of oxalyl-AMP as the intermediate in the synthesis of ODAP by LsAAE3.
a Extracted ion chromatograms of incubations of 100 mM Tris pH 8.0, 2 mM DTT, 5 mM ATP, 10 mM MgCl2, 0.3 mM sodium oxalate, 0.1 µM CoA with no enzyme, with LsAAE3, with LsAAE3 and desCoA replacing CoA, with LsAAE3, CoA and DAP. Black arrows indicate peak (m/z 418.0406) for oxalyl-AMP present in reactions with LsAAE3 and CoA or desCoA, which is absent when DAP is included as the oxalyl acceptor in the reaction. b Accurate mass determination with selected ion monitoring for peak m/z 418.0406. The calculated m/z was 418.04055, the observed was 418.0404 (indicated by black arrow) with an error of −0.36 ppm. c MS2 of selected ion m/z 418.0406. The calculated m/z for AMP in negative ion mode is 346.05581, the observed was 346.0551 with an error of −2.05 ppm. Source data are provided as a Source Data file.

Similar articles

References

    1. Goldsmith, M. et al. Identification and characterization of the key enzyme in the biosynthesis of the neurotoxin β-ODAP in grass pea. J. Biol. Chem.10.1016/j.jbc.2022.101806 (2022). - PMC - PubMed
    1. Edwards, A. et al. Genomics and biochemical analyses reveal a metabolon key to β-L-ODAP biosynthesis in Lathyrus sativus. Nat. Commun.14, 876 (2023). 10.1038/s41467-023-36503-2 - DOI - PMC - PubMed
    1. Emmrich, P. M. F. Genetic Improvement of Grass Pea (Lathyrus sativus) for Low β-L-ODAP Content. https://ueaeprints.uea.ac.uk/id/eprint/63944/ (2017).
    1. Quayle, J. R. Chemical synthesis of oxalyl-coenzyme A and its enzymic reduction to glyoxylate. Biochim Biophys. Acta57, 398–400 (1962). 10.1016/0006-3002(62)91142-3 - DOI - PubMed
    1. Leonardi, R., Zhang, Y.-M., Rock, C. O. & Jackowski, S. Coenzyme A: back in action. Prog. Lipid Res.44, 125–153 (2005). 10.1016/j.plipres.2005.04.001 - DOI - PubMed

MeSH terms

LinkOut - more resources