Protocol for engineering and validating a synthetic mitochondrial intermembrane bridge in mammalian cells

Gunjan Purohit, Martonio Ponte Viana, Oleh Khalimonchuk

Research output: Contribution to journalArticlepeer-review

Abstract

Membrane contact sites are recognized as critical means of intercompartmental communication. Here, we describe a protocol for engineering and validating a synthetic bridge between the inner and outer mitochondrial membranes to support functioning of the endogenous mitochondrial contact site and cristae organizing system (MICOS). A chimeric protein, MitoT, is stably expressed in cultured mammalian cells to bridge the mitochondrial membranes. This approach can be a valuable tool to study the function of the MICOS complex and associated proteins. For complete details on the use and execution of this protocol, please refer to Viana et al. (2021).

Original languageEnglish (US)
Article number101454
JournalSTAR Protocols
Volume3
Issue number3
DOIs
StatePublished - Sep 16 2022

Keywords

  • Biotechnology and bioengineering
  • Cell Biology
  • Cell Membrane
  • Cell culture
  • Cell isolation
  • Flow Cytometry/Mass Cytometry
  • Metabolism
  • Microscopy
  • Molecular Biology

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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