Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria

Research output: Contribution to journalArticleResearchpeer review

Authors

  • Yao Zheng
  • Cécile Cabassa-Hourton
  • Holger Eubel
  • Guillaume Chevreux
  • Laurent Lignieres
  • Emilie Crilat
  • Hans-Peter Braun
  • Sandrine Lebreton
  • Arnould Savouré

Research Organisations

External Research Organisations

  • Institute of Ecology and Environmental Sciences of Paris (iEES)
  • Université Paris Cité
  • Sorbonne Université
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Details

Original languageEnglish
Pages (from-to)917-934
Number of pages18
JournalJournal of experimental botany
Volume75
Issue number3
Early online date16 Oct 2023
Publication statusPublished - 2 Feb 2024

Abstract

Proline dehydrogenase (ProDH) and pyrroline-5-carboxylate (P5C) dehydrogenase (P5CDH) catalyze the oxidation of proline into glutamate via the intermediates P5C and glutamate-semialdehyde (GSA), which spontaneously interconvert. P5C and GSA are also intermediates in the production of glutamate from ornithine and α-ketoglutarate catalyzed by ornithine δ-aminotransferase (OAT). ProDH and P5CDH form a fused bifunctional PutA enzyme in Gram-negative bacteria and are associated in a bifunctional substrate channelling complex in Thermus thermophilus, but the physical proximity of ProDH and P5CDH in eukaryotes has not been described. Here we report evidence of physical proximity and interactions between Arabidopsis ProDH, P5CDH and OAT in the mitochondria of plants during dark-induced leaf senescence when all three enzymes are expressed. Pairwise interactions and localization of the three enzymes were investigated using bimolecular fluorescence complementation (BiFC) with confocal microscopy in tobacco and sub-mitochondrial fractionation in Arabidopsis. Evidence for a complex composed of ProDH, P5CDH, and OAT was revealed by co-migration of the proteins in native conditions upon gel electrophoresis. Co-immunoprecipitation coupled with mass spectrometry analysis confirmed the presence of the P5C metabolism complex in Arabidopsis. Pull-down assays further demonstrated a direct interaction between ProDH1 and P5CDH. P5C metabolism complexes may channel P5C among the constituent enzymes and directly provide electrons to the respiratory electron chain via ProDH.

Keywords

    Arabidopsis thaliana, mitochondria, ornithine δ-aminotransferase, proline catabolism, proline dehydrogenase, protein complex, pyrroline-5-carboxylate dehydrogenase

ASJC Scopus subject areas

Cite this

Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria. / Zheng, Yao; Cabassa-Hourton, Cécile; Eubel, Holger et al.
In: Journal of experimental botany, Vol. 75, No. 3, 02.02.2024, p. 917-934.

Research output: Contribution to journalArticleResearchpeer review

Zheng, Y, Cabassa-Hourton, C, Eubel, H, Chevreux, G, Lignieres, L, Crilat, E, Braun, H-P, Lebreton, S & Savouré, A 2024, 'Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria', Journal of experimental botany, vol. 75, no. 3, pp. 917-934. https://doi.org/10.1093/jxb/erad406
Zheng, Y., Cabassa-Hourton, C., Eubel, H., Chevreux, G., Lignieres, L., Crilat, E., Braun, H.-P., Lebreton, S., & Savouré, A. (2024). Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria. Journal of experimental botany, 75(3), 917-934. https://doi.org/10.1093/jxb/erad406
Zheng Y, Cabassa-Hourton C, Eubel H, Chevreux G, Lignieres L, Crilat E et al. Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria. Journal of experimental botany. 2024 Feb 2;75(3):917-934. Epub 2023 Oct 16. doi: 10.1093/jxb/erad406
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title = "Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria",
abstract = "Proline dehydrogenase (ProDH) and pyrroline-5-carboxylate (P5C) dehydrogenase (P5CDH) catalyze the oxidation of proline into glutamate via the intermediates P5C and glutamate-semialdehyde (GSA), which spontaneously interconvert. P5C and GSA are also intermediates in the production of glutamate from ornithine and α-ketoglutarate catalyzed by ornithine δ-aminotransferase (OAT). ProDH and P5CDH form a fused bifunctional PutA enzyme in Gram-negative bacteria and are associated in a bifunctional substrate channelling complex in Thermus thermophilus, but the physical proximity of ProDH and P5CDH in eukaryotes has not been described. Here we report evidence of physical proximity and interactions between Arabidopsis ProDH, P5CDH and OAT in the mitochondria of plants during dark-induced leaf senescence when all three enzymes are expressed. Pairwise interactions and localization of the three enzymes were investigated using bimolecular fluorescence complementation (BiFC) with confocal microscopy in tobacco and sub-mitochondrial fractionation in Arabidopsis. Evidence for a complex composed of ProDH, P5CDH, and OAT was revealed by co-migration of the proteins in native conditions upon gel electrophoresis. Co-immunoprecipitation coupled with mass spectrometry analysis confirmed the presence of the P5C metabolism complex in Arabidopsis. Pull-down assays further demonstrated a direct interaction between ProDH1 and P5CDH. P5C metabolism complexes may channel P5C among the constituent enzymes and directly provide electrons to the respiratory electron chain via ProDH.",
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note = "Funding Information: This work was supported by the Hubert Curien PROCOPE program between Germany and France (grant no. 46711VJ) funded by the Ministry of Foreign Affairs and by Sorbonne University. This work was also supported by a grant from the China Scholarship Council to YZ. We thank R{\'e}gion Ile de France for financial support of the ProteoSeine@IJM platform.",
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TY - JOUR

T1 - Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria

AU - Zheng, Yao

AU - Cabassa-Hourton, Cécile

AU - Eubel, Holger

AU - Chevreux, Guillaume

AU - Lignieres, Laurent

AU - Crilat, Emilie

AU - Braun, Hans-Peter

AU - Lebreton, Sandrine

AU - Savouré, Arnould

N1 - Funding Information: This work was supported by the Hubert Curien PROCOPE program between Germany and France (grant no. 46711VJ) funded by the Ministry of Foreign Affairs and by Sorbonne University. This work was also supported by a grant from the China Scholarship Council to YZ. We thank Région Ile de France for financial support of the ProteoSeine@IJM platform.

PY - 2024/2/2

Y1 - 2024/2/2

N2 - Proline dehydrogenase (ProDH) and pyrroline-5-carboxylate (P5C) dehydrogenase (P5CDH) catalyze the oxidation of proline into glutamate via the intermediates P5C and glutamate-semialdehyde (GSA), which spontaneously interconvert. P5C and GSA are also intermediates in the production of glutamate from ornithine and α-ketoglutarate catalyzed by ornithine δ-aminotransferase (OAT). ProDH and P5CDH form a fused bifunctional PutA enzyme in Gram-negative bacteria and are associated in a bifunctional substrate channelling complex in Thermus thermophilus, but the physical proximity of ProDH and P5CDH in eukaryotes has not been described. Here we report evidence of physical proximity and interactions between Arabidopsis ProDH, P5CDH and OAT in the mitochondria of plants during dark-induced leaf senescence when all three enzymes are expressed. Pairwise interactions and localization of the three enzymes were investigated using bimolecular fluorescence complementation (BiFC) with confocal microscopy in tobacco and sub-mitochondrial fractionation in Arabidopsis. Evidence for a complex composed of ProDH, P5CDH, and OAT was revealed by co-migration of the proteins in native conditions upon gel electrophoresis. Co-immunoprecipitation coupled with mass spectrometry analysis confirmed the presence of the P5C metabolism complex in Arabidopsis. Pull-down assays further demonstrated a direct interaction between ProDH1 and P5CDH. P5C metabolism complexes may channel P5C among the constituent enzymes and directly provide electrons to the respiratory electron chain via ProDH.

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KW - ornithine δ-aminotransferase

KW - proline catabolism

KW - proline dehydrogenase

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DO - 10.1093/jxb/erad406

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