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

Gunjan Purohit; Martonio Ponte Viana; Oleh Khalimonchuk

doi:10.1016/j.xpro.2022.101454

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

Gunjan Purohit, Martonio Ponte Viana, Oleh Khalimonchuk

Biochemistry

Research output: Contribution to journal › Article › peer-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 language	English (US)
Article number	101454
Journal	STAR Protocols
Volume	3
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2022.101454
State	Published - 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

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2022.101454

Cite this

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title = "Protocol for engineering and validating a synthetic mitochondrial intermembrane bridge in mammalian cells",

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).",

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AU - Purohit, Gunjan

AU - Viana, Martonio Ponte

AU - Khalimonchuk, Oleh

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AB - 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).

KW - Biotechnology and bioengineering

KW - Cell Biology

KW - Cell Membrane

KW - Cell culture

KW - Cell isolation

KW - Flow Cytometry/Mass Cytometry

KW - Metabolism

KW - Microscopy

KW - Molecular Biology

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ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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