Proteomic and functional analysis of proline dehydrogenase 1 link proline catabolism to mitochondrial electron transport in Arabidopsis thaliana.
Proline accumulates in many plant species in response to environmental stresses. Upon relief from stress, proline is rapidly oxidized in mitochondria by proline dehydrogenase (ProDH) and then by pyrroline-5-carboxylate dehydrogenase (P5CDH). Two ProDH genes have been identified in the genome of the model plant Arabidopsis thaliana To gain a better understanding of ProDH1 functions in mitochondria, proteomic analysis was performed. ProDH1 polypeptides were identified in Arabidopsis mitochondria by immunoblotting gels after 2D blue native (BN)-SDS/PAGE, probing them with an anti-ProDH antibody and analysing protein spots by MS. The 2D gels showed that ProDH1 forms part of a low-molecular-mass (70-140 kDa) complex in the mitochondrial membrane. To evaluate the contribution of each isoform to proline oxidation, mitochondria were isolated from wild-type (WT) and prodh1, prodh2, prodh1prodh2 and p5cdh mutants. ProDH activity was high for genotypes in which ProDH, most likely ProDH1, was strongly induced by proline. Respiratory measurements indicate that ProDH1 has a role in oxidizing excess proline and transferring electrons to the respiratory chain.
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Cécile
C
Cabassa-Hourton
Institute of Plant Genetics, Plant Proteomics, Leibniz University Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany.
Peter
P
Schertl
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Marianne
M
Bordenave-Jacquemin
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France Laboratoire des Plantes Extrêmophiles, Centre de Biotechnologie de Borj-Cedria (CBBC), BP 901, Hammam-Lif 2050, Tunisia.
Kaouthar
K
Saadallah
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Anne
A
Guivarc'h
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Sandrine
S
Lebreton
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Séverine
S
Planchais
Institute of Plant Genetics, Plant Proteomics, Leibniz University Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany.
Jennifer
J
Klodmann
Institute of Plant Genetics, Plant Proteomics, Leibniz University Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany.
Holger
H
Eubel
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Emilie
E
Crilat
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Delphine
D
Lefebvre-De Vos
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Thanos
T
Ghelis
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Luc
L
Richard
Laboratoire des Plantes Extrêmophiles, Centre de Biotechnologie de Borj-Cedria (CBBC), BP 901, Hammam-Lif 2050, Tunisia.
Chedly
C
Abdelly
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France.
Pierre
P
Carol
Institute of Plant Genetics, Plant Proteomics, Leibniz University Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany braun@genetik.uni-hannover.de arnould.savoure@upmc.fr.
Hans-Peter
HP
Braun
Sorbonne Universités, UPMC Univ Paris 06, iEES, UMR 7618, UPMC Paris 06-Sorbonne (UPEC, UPMC, CNRS, IRD, INRA, Paris Diderot), case 237, 4 place Jussieu, F-75252 Paris cedex 05, France braun@genetik.uni-hannover.de arnould.savoure@upmc.fr.
Arnould
A
Savouré
D017360:Arabidopsis / Q000201:enzymology / Q000378:metabolism*; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry; D008928:Mitochondria / Q000378:metabolism*; D011392:Proline / Q000378:metabolism*; D011394:Proline Oxidase / Q000378:metabolism*; D020543:Proteome
D020543:Proteome; D011392:Proline; D011394:Proline Oxidase
Arabidopsis thaliana; electron transfer chain; mitochondria; proline; proline dehydrogenase (ProDH)