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  Probing the environment of emerin by Enhanced ascorbate peroxidase 2 (APEX2)-mediated proximity labeling.

Müller, M., James, C., Lenz, C., Urlaub, H., & Kehlenbach, R. H. (2020). Probing the environment of emerin by Enhanced ascorbate peroxidase 2 (APEX2)-mediated proximity labeling. Cells, 9(3): 605. doi:10.3390/cells9030605.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0005-DA20-1 Version Permalink: http://hdl.handle.net/21.11116/0000-0006-D13A-D
Genre: Journal Article

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 Creators:
Müller, M., Author
James, C., Author
Lenz, C.1, Author              
Urlaub, H.1, Author              
Kehlenbach, R. H., Author
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1Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society, ou_578613              

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Free keywords: proximity labeling; APEX; RAPIDS; emerin; inner nuclear membrane
 Abstract: Emerin is one of the best characterized proteins of the inner nuclear membrane, but can also occur at the level of the endoplasmic reticulum. We now use enhanced ascorbate peroxidase 2 (APEX2) to probe the environment of emerin. APEX2 can be used as a genetic tag that produces short-lived yet highly reactive biotin species, allowing the modification of proteins that interact with or are in very close proximity to the tagged protein. Biotinylated proteins can be isolated using immobilized streptavidin and analyzed by mass spectrometry. As an alternative to the standard approach with a genetic fusion of APEX2 to emerin, we also used RAPIDS (rapamycin- and APEX-dependent identification of proteins by SILAC), a method with improved specificity, where the peroxidase interacts with the protein of interest (i.e., emerin) only upon addition of rapamycin to the cells. We compare these different approaches, which, together, identify well-known interaction partners of emerin like lamin A and the lamina associated polypeptide 1 (LAP1), as well as novel proximity partners.

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Language(s): eng - English
 Dates: 2020-03-03
 Publication Status: Published online
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 Rev. Type: Peer
 Identifiers: DOI: 10.3390/cells9030605
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Title: Cells
Source Genre: Journal
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Pages: 18 Volume / Issue: 9 (3) Sequence Number: 605 Start / End Page: - Identifier: -