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Proteomic mapping by rapamycin-dependent targeting of APEX2 identifies binding partners of VAPB at the inner nuclear membrane

James, Christina; Müller, Marret; Goldberg, Martin W.; Lenz, Christof; Urlaub, Henning; Kehlenbach, Ralph H.

Proteomic mapping by rapamycin-dependent targeting of APEX2 identifies binding partners of VAPB at the inner nuclear membrane Thumbnail


Authors

Christina James

Marret Müller

Christof Lenz

Henning Urlaub

Ralph H. Kehlenbach



Abstract

Vesicle-associated membrane protein–associated protein B (VAPB) is a tail-anchored protein that is present at several contact sites of the endoplasmic reticulum (ER). We now show by immunoelectron microscopy that VAPB also localizes to the inner nuclear membrane (INM). Using a modified enhanced ascorbate peroxidase 2 (APEX2) approach with rapamycin-dependent targeting of the peroxidase to a protein of interest, we searched for proteins that are in close proximity to VAPB, particularly at the INM. In combination with stable isotope labeling with amino acids in cell culture (SILAC), we confirmed many well-known interaction partners at the level of the ER with a clear distinction between specific and nonspecific hits. Furthermore, we identified emerin, TMEM43, and ELYS as potential interaction partners of VAPB at the INM and the nuclear pore complex, respectively.

Citation

James, C., Müller, M., Goldberg, M. W., Lenz, C., Urlaub, H., & Kehlenbach, R. H. (2019). Proteomic mapping by rapamycin-dependent targeting of APEX2 identifies binding partners of VAPB at the inner nuclear membrane. Journal of Biological Chemistry, 294(44), 16241-16254. https://doi.org/10.1074/jbc.ra118.007283

Journal Article Type Article
Online Publication Date Sep 13, 2019
Publication Date Nov 1, 2019
Deposit Date Sep 25, 2019
Publicly Available Date Sep 13, 2020
Journal Journal of Biological Chemistry
Print ISSN 0021-9258
Electronic ISSN 1083-351X
Publisher American Society for Biochemistry and Molecular Biology
Peer Reviewed Peer Reviewed
Volume 294
Issue 44
Pages 16241-16254
DOI https://doi.org/10.1074/jbc.ra118.007283

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Copyright Statement
This research was originally published in James, Christina, Müller, Marret, Goldberg, Martin W., Lenz, Christof, Urlaub, Henning & Kehlenbach, Ralph H. (2019). Proteomic mapping by rapamycin-dependent targeting of APEX2 identifies binding partners of VAPB at the inner nuclear membrane. Journal of Biological Chemistry 294(44): 16241-16254 © the American Society for Biochemistry and Molecular Biology.







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