A quantitative fluorescence‐based approach to study mitochondrial protein import

2023-03-20 | journal article. A publication with affiliation to the University of Göttingen.

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​A quantitative fluorescence‐based approach to study mitochondrial protein import​
Jain, N.; Gomkale, R.; Bernhard, O.; Rehling, P. & Cruz‐Zaragoza, L. D.​ (2023) 
EMBO Reports24(5).​ DOI: https://doi.org/10.15252/embr.202255760 

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Authors
Jain, Naintara; Gomkale, Ridhima; Bernhard, Olaf; Rehling, Peter; Cruz‐Zaragoza, Luis Daniel
Abstract
Mitochondria play central roles in cellular energy production and metabolism. Most proteins required to carry out these functions are synthesized in the cytosol and imported into mitochondria. A growing number of metabolic disorders arising from mitochondrial dysfunction can be traced to errors in mitochondrial protein import. The mechanisms underlying the import of precursor proteins are commonly studied using radioactively labeled precursor proteins imported into purified mitochondria. Here, we establish a fluorescence-based import assay to analyze protein import into mitochondria. We show that fluorescently labeled precursors enable import analysis with similar sensitivity to those using radioactive precursors, yet they provide the advantage of quantifying import with picomole resolution. We adapted the import assay to a 96-well plate format allowing for fast analysis in a screening-compatible format. Moreover, we show that fluorescently labeled precursors can be used to monitor the assembly of the F1F0 ATP synthase in purified mitochondria. Thus, we provide a sensitive fluorescence-based import assay that enables quantitative and fast import analysis.
Issue Date
20-March-2023
Journal
EMBO Reports 
Project
EXC 2067: Multiscale Bioimaging 
SFB 1190: Transportmaschinen und Kontaktstellen zellulärer Kompartimente 
SFB 1190 | P13: Protein Transport über den mitochondrialen Carrier Transportweg 
Working Group
RG Rehling (Mitochondrial Protein Biogenesis) 
External URL
https://mbexc.uni-goettingen.de/literature/publications/668
https://sfb1190.med.uni-goettingen.de/production/literature/publications/217
ISSN
1469-221X; 1469-3178
eISSN
1469-3178
Language
English
Sponsor
Deutsche Forschungsgemeinschaft (DFG) http://dx.doi.org/10.13039/501100001659

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