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

Authors

Gunjan Purohit, University of Nebraska - LincolnFollow
Martonio Ponte Viana, University of Nebraska -Lincoln
Oleh Khalimonchuk, University of Nebraska-LincolnFollow

Date of this Version

9-16-2022

Document Type

Article

Citation

Purohit et al., STAR Protocols 3, 101454 September 16, 2022. https://doi.org/10.1016/ j.xpro.2022.101454

Comments

This is an open access article under the CC BY-NC-ND license

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 themitochondrial membranes. This approach can be a valuable tool to study the function of the MICOS complex and associated proteins.