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  Protein Import Assay into Mitochondria Isolated from Human Cells

Murschall, L. M., Peker, E., MacVicar, T., Langer, T., & Riemer, J. (2021). Protein Import Assay into Mitochondria Isolated from Human Cells. Bio Protoc, 11(12), e4057. doi:10.21769/BioProtoc.4057.

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Murschall, L. M., Author
Peker, E., Author
MacVicar, T.1, Author           
Langer, T.1, Author           
Riemer, J., Author
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1Department Langer - Mitochondrial Proteostasis, Max Planck Institute for Biology of Ageing, Max Planck Society, ou_3393994              

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Free keywords: Cell-free protein synthesis Human tissue culture cells In organello import Isolated mitochondria Radiolabeled proteins
 Abstract: Mitochondria are essential organelles containing approximately 1,500 proteins. Only approximately 1% of these proteins are synthesized inside mitochondria, whereas the remaining 99% are synthesized as precursors on cytosolic ribosomes and imported into the organelle. Various tools and techniques to analyze the import process have been developed. Among them, in vitro reconstituted import systems are of importance to study these processes in detail. These experiments monitor the import reaction of mitochondrial precursors that were previously radiolabeled in a cell-free environment. However, the methods described have been mostly performed in mitochondria isolated from S. cerevisiae. Here, we describe the adaptation of this powerful assay to import proteins into crude mitochondria isolated from human tissue culture cells. Graphic abstract: Overview of the assay to monitor protein import into mitochondria isolated from human cells.

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 Dates: 2021-06-202021-06-20
 Publication Status: Issued
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 Identifiers: Other: 34263000
DOI: 10.21769/BioProtoc.4057
ISSN: 2331-8325 (Electronic)2331-8325 (Linking)
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Title: Bio Protoc
Source Genre: Journal
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Pages: - Volume / Issue: 11 (12) Sequence Number: - Start / End Page: e4057 Identifier: -