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Plastids pp 173-188 | Cite as

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

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

Abstract

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 

Notes

Acknowledgments

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.

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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

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