Plastids pp 173-188 | Cite as

Analysis of the MTL Supercomplex at Contact Sites Between Mitochondria and Plastids

  • Morgane MichaudEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1829)


Plastids are organelles playing fundamental roles in different cellular processes such as energy metabolism or lipid biosynthesis. To fulfill their biogenesis and their function in the cell, plastids have to communicate with other cellular compartments. This communication can be mediated by the establishment of direct contact sites between plastids envelop and other organelles. These contacts are dynamic structures that are modified in response to stress. As example, during phosphate (Pi) starvation, the number of contact sites between plastids and mitochondria significantly increases. In this situation, these contacts play an important role in the transfer of galactoglycerolipids from plastids to mitochondria. Recently, Pi starvation stress was used to identify key proteins involved in the traffic of galactoglycerolipids from plastids to mitochondria in Arabidopsis thaliana. A mitochondrial lipoprotein complex called MTL (mitochondrial transmembrane lipoprotein complex) was identified. This complex contains mitochondrial proteins but also proteins located in the plastid envelope, suggesting its presence at the plastid–mitochondria junction. This chapter describes the protocol to isolate the MTL complex by clear-native polyacrylamide gel electrophoresis (CN-PAGE) from the mitochondrial fraction of Arabidopsis cell cultures and the methods to study different features of this complex.

Key words

Plastids Mitochondria MTL complex Membrane contact sites Arabidopsis thaliana Phosphate starvation CN-PAGE 



This work was supported by the Intramural Research Program of the National Institute of Diabetes and Digestive and Kidney Diseases. I thank Catherine Albrieux and Joan Guldemann for critical reading of the manuscript.


  1. 1.
    Boudière L, Michaud M, Petroutsos D et al (2014) Glycerolipids in photosynthesis: composition, synthesis and trafficking. Biochim Biophys Acta 1837(4):470–480. Scholar
  2. 2.
    Li-Beisson Y, Shorrosh B, Beisson F et al (2013) Acyl-lipids metabolism. Arabidopsis Book 11:e0161. Scholar
  3. 3.
    Boudiere L, Botte CY, Saidani N et al (2012) Galvestine-1, a novel chemical probe for the study of the glycerolipid homeostasis system in plant cells. Mol BioSyst 8(8):2023–2035., 2014. Scholar
  4. 4.
    Raghothama KG (1999) Phosphate acquisition. Annu Rev Plant Physiol Plant Mol Biol 50:665–693. Scholar
  5. 5.
    Zhang Z, Liao H, Lucas WJ (2014) Molecular mechanisms underlying phosphate sensing, signaling, and adaptation in plants. J Integr Plant Biol 56(3):192–220. Scholar
  6. 6.
    Poirier Y, Thoma S, Somerville C et al (1991) Mutant of Arabidopsis deficient in xylem loading of phosphate. Plant Physiol 97(3):1087–1093CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Jouhet J, Marechal E, Bligny R et al (2003) Transient increase of phosphatidylcholine in plant cells in response to phosphate deprivation. FEBS Lett 544(1–3):63–68. Scholar
  8. 8.
    Andersson MX, Stridh MH, Larsson KE et al (2003) Phosphate-deficient oat replaces a major portion of the plasma membrane phospholipids with the galactolipid digalactosyldiacylglycerol. FEBS Lett 537(1–3):128–132CrossRefPubMedGoogle Scholar
  9. 9.
    Jouhet J, Marechal E, Baldan B et al (2004) Phosphate deprivation induces transfer of DGDG galactolipid from chloroplast to mitochondria. J Cell Biol 167(5):863–874. Scholar
  10. 10.
    Andersson MX, Larsson KE, Tjellstrom H et al (2005) Phosphate-limited oat. The plasma membrane and the tonoplast as major targets for phospholipid-to-glycolipid replacement and stimulation of phospholipases in the plasma membrane. J Biol Chem 280(30):27578–27586. Scholar
  11. 11.
    Michaud M, Gros V, Tardif M et al (2016) AtMic60 is involved in plant mitochondria lipid trafficking and is part of a large complex. Curr Biol 26:627–639. Scholar
  12. 12.
    Michaud M, Prinz WA, Jouhet J (2017) Glycerolipid synthesis and lipid trafficking in plant mitochondria. FEBS J 284:376–390. Scholar
  13. 13.
    von der Malsburg K, Muller JM, Bohnert M et al (2011) Dual role of mitofilin in mitochondrial membrane organization and protein biogenesis. Dev Cell 21(4):694–707. Scholar
  14. 14.
    Axelos M, Curie C, Mazzolini L et al (1992) A protocol for transient gene expression in Arabidopsis thaliana protoplasts isolated from cell suspension cultures. Plant Physiol Biochem 30:123–128Google Scholar
  15. 15.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  16. 16.
    Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37(8):911–917CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney DiseasesNIHBethesdaUSA
  2. 2.Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Institut National de la Recherche Agronomique, Commissariat à l’Energie Atomique et aux Energies Alternatives, CEA GrenobleUMR5168, Université Grenoble AlpesGrenobleFrance

Personalised recommendations