MERLIN: a novel BRET-based proximity biosensor for studying mitochondria–ER contact sites

DOI Web Site 被引用文献3件
  • Vanessa Hertlein
    Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany
  • Hector Flores-Romero
    Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany
  • Kushal K Das
    Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany
  • Sebastian Fischer
    University of Heidelberg, Heidelberg, Germany
  • Michael Heunemann
    Center for Plant Molecular Biology, University of Tübingen, Tübingen, Germany
  • Maria Calleja-Felipe
    Molecular Cognition Laboratory, Biophysics Institute, Consejo Superior de Investigaciones Cientificas, University of the Basque Country (UPV)/Euskal Herriko University, Campus Universidad del País Vasco, Leioa, Spain
  • Shira Knafo
    Molecular Cognition Laboratory, Biophysics Institute, Consejo Superior de Investigaciones Cientificas, University of the Basque Country (UPV)/Euskal Herriko University, Campus Universidad del País Vasco, Leioa, Spain
  • Katharina Hipp
    Max Planck Institute for Developmental Biology, Tübingen, Germany
  • Klaus Harter
    Center for Plant Molecular Biology, University of Tübingen, Tübingen, Germany
  • Julia C Fitzgerald
    Hertie-Institute for Clinical Brain Research, University of Tübingen and German Centre for Neurodegenerative Diseases (DZNE), Tübingen, Germany
  • Ana J García-Sáez
    Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany

抄録

<jats:p>The contacts between the ER and mitochondria play a key role in cellular functions such as the exchange of lipids and calcium between both organelles, as well as in apoptosis and autophagy signaling. The molecular architecture and spatiotemporal regulation of these distinct contact regions remain obscure and there is a need for new tools that enable tackling these questions. Here, we present a new bioluminescence resonance energy transfer–based biosensor for the quantitative analysis of distances between the ER and mitochondria that we call MERLIN (Mitochondria–ER Length Indicator Nanosensor). The main advantages of MERLIN compared with available alternatives are that it does not rely on the formation of artificial physical links between the two organelles, which could lead to artifacts, and that it allows to study contact site reversibility and dynamics. We show the applicability of MERLIN by characterizing the role of the mitochondrial dynamics machinery on the contacts of this organelle with the ER.</jats:p>

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