Lipoic acid-dependent oxidative catabolism of α-keto acids in mitochondria provides evidence for branched-chain amino acid catabolism in Arabidopsis

被引：147

作者：

Taylor, NL ^{[1
]}

Heazlewood, JL ^{[1
]}

Day, DA ^{[1
]}

Millar, AH ^{[1
]}

机构：

[1] Univ Western Australia, Sch Biomed & Chem Sci, Plant Mol Biol Grp, Crawley, WA 6009, Australia

来源：

PLANT PHYSIOLOGY | 2004年 / 134卷 / 02期

关键词：

D O I：

10.1104/pp.103.035675

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

Lipoic acid-dependent pathways of alpha-keto acid oxidation by mitochondria were investigated in pea (Pisum sativum), rice (Oryza sativa), and Arabidopsis. Proteins containing covalently bound lipoic acid were identified on isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis separations of mitochondrial proteins by the use of antibodies raised to this cofactor. All these proteins were identified by tandem mass spectrometry. Lipoic acid-containing acyltransferases from pyruvate dehydrogenase complex and alpha-ketoglutarate dehydrogenase complex were identified from all three species. In addition, acyltransferases from the branched-chain dehydrogenase complex were identified in both Arabidopsis and rice mitochondria. The substrate-dependent reduction of NAD(+) was analyzed by spectrophotometry using specific alpha-keto acids. Pyruvate- and alpha-ketoglutarate-dependent reactions were measured in all three species. Activity of the branched-chain dehydrogenase complex was only measurable in Arabidopsis mitochondria using substrates that represented the alpha-keto acids derived by deamination of branched-chain amino acids (Val [valine], leucine, and isoleucine). The rate of branched-chain amino acid- and alpha-keto acid-dependent oxygen consumption by intact Arabidopsis mitochondria was highest with Val and the Val-derived alpha-keto acid, alpha-ketoisovaleric acid. Sequencing of peptides derived from trypsination of Arabidopsis mitochondrial proteins revealed the presence of many of the enzymes required for the oxidation of all three branched-chain amino acids. The potential role of branched-chain amino acid catabolism as an oxidative phosphorylation energy source or as a detoxification pathway during plant stress is discussed.

引用

页码：838 / 848

页数：11

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