Chapter 13 Localization and function of the 2Fe-2S outer mitochondrial membrane protein mitoNEET
Related Experiment Video
Updated: Jun 24, 2026

An Improved Method to Isolate Mitochondrial Contact Sites
Published on: June 16, 2023
Sandra E Wiley1, Matthew J Rardin, Jack E Dixon
1Department of Pharmacology, University of California, San Diego, La Jolla, California, USA.
Abstract:
MitoNEET is an integral protein of the outer mitochondrial membrane and is the flagship of a small family of proteins whose hallmark is the presence of a CDGSH domain. Initially annotated as a zinc finger, the CDGSH domain actually binds a redox-active 2Fe-2S cluster, giving mitoNEET the distinction of being the first 2Fe-2S protein identified in the outer membrane of mitochondria. This chapter describes methods for isolating mitochondrial membrane fractions that are enriched in mitoNEET, generating constructs for the expression of recombinant mitoNEET protein and analyzing the 2Fe-2S cluster of mitoNEET in vitro.
Related Concept Videos
Mitochondrial Protein Sorting
Most of these mitochondrial proteins are encoded by the nucleus and imported to the mitochondria as unfolded or loosely folded precursors. Mitochondrial precursors...
Protein Transport into the Inner Mitochondrial Membrane
Transport of mitochondrial precursors across the TIM23 channel is driven by...
Translocation of Proteins into the Mitochondria
Sorting of outer membrane proteins:
Mitochondrial outer membrane proteins are of two types: the transmembrane, beta-barrel porins, and the membrane-anchored, alpha-helical proteins. Beta-barrel porin precursors are translocated by the TOM complex and inserted into the outer mitochondrial membrane by the SAM complex. In contrast,...
Mitochondrial Membranes
Mitochondrial Membranes
The Inner Mitochondrial Membrane

