Protocol for real-time imaging of membrane fission by mitofissin.

STAR Protoc

Institute of Microbial Chemistry (BIKAKEN), Shinagawa-ku, Tokyo 141-0021, Japan; Institute for Genetic Medicine, Hokkaido University, Sapporo, Hokkaido 060-0815, Japan. Electronic address:

Published: December 2023


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Article Abstract

Yeast mitofissin Atg44 is a mitochondrial intermembrane space protein that causes membrane fission required for mitophagy. Here, we present a protocol for observing Atg44-mediated membrane fission. We describe steps for recombinant Atg44 purification, lipid nanotube preparation as model membranes, and Atg44-mediated membrane fission real-time observation. We then detail procedures for tube radius estimation using confocal microscopy. This protocol can also be adapted to the study of membrane fission by other proteins. For complete details on the use and execution of this protocol, please refer to Fukuda et al. (2023)..

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10520655PMC
http://dx.doi.org/10.1016/j.xpro.2023.102590DOI Listing

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