Enzymes of glycolysis are functionally associated with the mitochondrion in Arabidopsis cells

被引:296
作者
Giegé, P
Heazlewood, JL
Roessner-Tunali, U
Millar, AH
Fernie, AR
Leaver, CJ
Sweetlove, LJ
机构
[1] Univ Oxford, Dept Plant Sci, Oxford OX1 3RB, England
[2] Inst Biol Mol Plantes, F-67084 Strasbourg, France
[3] Univ Western Australia, Sch Biomed & Chem Sci, Crawley, WA 6009, Australia
[4] Max Planck Inst Mol Physiol, Dept Lothar Willmitzer, D-14476 Golm, Germany
关键词
D O I
10.1105/tpc.012500
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Mitochondria fulfill a wide range of metabolic functions in addition to the synthesis of ATP and contain a diverse array of proteins to perform these functions. Here, we present the unexpected discovery of the presence of the enzymes of glycolysis in a mitochondrial fraction of Arabidopsis cells. Proteomic analyses of this mitochondrial fraction revealed the presence of 7 of the 10 enzymes that constitute the glycolytic pathway. Four of these enzymes (glyceraldehyde-3-P dehydrogenase, aldolase, phosphoglycerate mutase, and enolase) were also identified in an intermembrane space/outer mitochondrial membrane fraction. Enzyme activity assays confirmed that the entire glycolytic pathway was present in preparations of isolated Arabidopsis mitochondria, and the sensitivity of these activities to protease treatments indicated that the glycolytic enzymes are present on the outside of the mitochondrion. The association of glycolytic enzymes with mitochondria was confirmed in vivo by the expression of enolase- and aldolase-yellow fluorescent protein fusions in Arabidopsis protoplasts. The yellow fluorescent protein fluorescence signal showed that these two fusion proteins are present throughout the cytosol but are also concentrated in punctate regions that colocalized with the mitochondrion-specific probe Mitotracker Red. Furthermore, when supplied with appropriate cofactors, isolated, intact mitochondria were capable of the metabolism of C-13-glucose to C-13-labeled intermediates of the trichloroacetic acid cycle, suggesting that the complete glycolytic sequence is present and active in this subcellular fraction. On the basis of these data, we propose that the entire glycolytic pathway is associated with plant mitochondria by attachment to the cytosolic face of the outer mitochondrial membrane and that this microcompartmentation of glycolysis allows pyruvate to be provided directly to the mitochondrion, where it is used as a respiratory substrate.
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页码:2140 / 2151
页数:12
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