title: Lipoate-Protein Ligase and Octanoyltransferase Are Essential for Protein Lipoylation in Mitochondria of Arabidopsis.
abstract: Prosthetic lipoyl groups are required for the function of several essential multienzyme complexes, such as pyruvate dehydrogenase (PDH), α-ketoglutarate dehydrogenase (KGDH), and the glycine cleavage system (glycine decarboxylase [GDC]). How these proteins are lipoylated has been extensively studied in prokaryotes and yeast (Saccharomyces cerevisiae), but little is known for plants. We earlier reported that mitochondrial fatty acid synthesis by ketoacyl-acyl carrier protein synthase is not vital for protein lipoylation in Arabidopsis (Arabidopsis thaliana) and does not play a significant role in roots. Here, we identify Arabidopsis lipoate-protein ligase (AtLPLA) as an essential mitochondrial enzyme that uses octanoyl-nucleoside monophosphate and possibly other donor substrates for the octanoylation of mitochondrial PDH-E2 and GDC H-protein; it shows no reactivity with bacterial and possibly plant KGDH-E2. The octanoate-activating enzyme is unknown, but we assume that it uses octanoyl moieties provided by mitochondrial β-oxidation. AtLPLA is essential for the octanoylation of PDH-E2, whereas GDC H-protein can optionally also be octanoylated by octanoyltransferase (LIP2) using octanoyl chains provided by mitochondrial ketoacyl-acyl carrier protein synthase to meet the high lipoate requirement of leaf mesophyll mitochondria. Similar to protein lipoylation in yeast, LIP2 likely also transfers octanoyl groups attached to the H-protein to KGDH-E2 but not to PDH-E2, which is exclusively octanoylated by LPLA. We suggest that LPLA and LIP2 together provide a basal protein lipoylation network to plants that is similar to that in other eukaryotes.
authors.affiliation: Department of Plant Physiology, University of Rostock, D-18059 Rostock, Germany (R.E., H.B.);Department of Plant Physiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany (C.H., E.N.); andMax-Planck-Institut für Molekulare Pflanzenphysiologie, 14476 Potsdam-Golm, Germany (A.F., A.R.F.).
authors.forename: Ralph
authors.initials: R
authors.lastname: Ewald
authors.affiliation: Department of Plant Physiology, University of Rostock, D-18059 Rostock, Germany (R.E., H.B.);Department of Plant Physiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany (C.H., E.N.); andMax-Planck-Institut für Molekulare Pflanzenphysiologie, 14476 Potsdam-Golm, Germany (A.F., A.R.F.).
authors.forename: Christiane
authors.initials: C
authors.lastname: Hoffmann
authors.affiliation: Department of Plant Physiology, University of Rostock, D-18059 Rostock, Germany (R.E., H.B.);Department of Plant Physiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany (C.H., E.N.); andMax-Planck-Institut für Molekulare Pflanzenphysiologie, 14476 Potsdam-Golm, Germany (A.F., A.R.F.).
authors.forename: Alexandra
authors.initials: A
authors.lastname: Florian
authors.affiliation: Department of Plant Physiology, University of Rostock, D-18059 Rostock, Germany (R.E., H.B.);Department of Plant Physiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany (C.H., E.N.); andMax-Planck-Institut für Molekulare Pflanzenphysiologie, 14476 Potsdam-Golm, Germany (A.F., A.R.F.).
authors.forename: Ekkehard
authors.initials: E
authors.lastname: Neuhaus
authors.affiliation: Department of Plant Physiology, University of Rostock, D-18059 Rostock, Germany (R.E., H.B.);Department of Plant Physiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany (C.H., E.N.); andMax-Planck-Institut für Molekulare Pflanzenphysiologie, 14476 Potsdam-Golm, Germany (A.F., A.R.F.).
authors.forename: Alisdair R
authors.initials: AR
authors.lastname: Fernie
authors.affiliation: Department of Plant Physiology, University of Rostock, D-18059 Rostock, Germany (R.E., H.B.);Department of Plant Physiology, University of Kaiserslautern, D-67663 Kaiserslautern, Germany (C.H., E.N.); andMax-Planck-Institut für Molekulare Pflanzenphysiologie, 14476 Potsdam-Golm, Germany (A.F., A.R.F.) hermann.bauwe@uni-rostock.de.
authors.forename: Hermann
authors.initials: H
authors.lastname: Bauwe