APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest
Deciphering the sub-compartmental location of a given protein of interest may help explain its physiological function, but it can be challenging to do using optical or biochemical methods. Imaging with electron microscopy (EM) can provide highly resolved mapping of proteins; however, EM requires com...
| Main Authors: | , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
Elsevier
2017
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| Online Access: | http://hdl.handle.net/1721.1/110551 https://orcid.org/0000-0002-8277-5226 |
| _version_ | 1826197136103964672 |
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| author | Lee, Song-Yi Kang, Myeong-Gyun Park, Jong-Seok Lee, Geunsik Rhee, Hyun-Woo Ting, Alice Y |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Lee, Song-Yi Kang, Myeong-Gyun Park, Jong-Seok Lee, Geunsik Rhee, Hyun-Woo Ting, Alice Y |
| author_sort | Lee, Song-Yi |
| collection | MIT |
| description | Deciphering the sub-compartmental location of a given protein of interest may help explain its physiological function, but it can be challenging to do using optical or biochemical methods. Imaging with electron microscopy (EM) can provide highly resolved mapping of proteins; however, EM requires complex sample preparation and a specialized facility. Here, we use engineered ascorbate peroxidase (APEX)-generated molecular labeling patterns to provide information regarding intracellular microenvironments in living cells. Using APEX labeling of specific proteins, we uncovered subcellular localization at sub-compartmental resolution and successfully elucidated the membrane protein topology of HMOX1 and sub-mitochondrial localization of recently identified mitochondrial proteins. This method can be expanded to confirm sub-mitochondrial localization and membrane topologies of previously identified mitochondrial proteins. |
| first_indexed | 2024-09-23T10:43:22Z |
| format | Article |
| id | mit-1721.1/110551 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:43:22Z |
| publishDate | 2017 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1105512022-09-30T22:31:23Z APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest Lee, Song-Yi Kang, Myeong-Gyun Park, Jong-Seok Lee, Geunsik Rhee, Hyun-Woo Ting, Alice Y Massachusetts Institute of Technology. Department of Chemistry Ting, Alice Y Deciphering the sub-compartmental location of a given protein of interest may help explain its physiological function, but it can be challenging to do using optical or biochemical methods. Imaging with electron microscopy (EM) can provide highly resolved mapping of proteins; however, EM requires complex sample preparation and a specialized facility. Here, we use engineered ascorbate peroxidase (APEX)-generated molecular labeling patterns to provide information regarding intracellular microenvironments in living cells. Using APEX labeling of specific proteins, we uncovered subcellular localization at sub-compartmental resolution and successfully elucidated the membrane protein topology of HMOX1 and sub-mitochondrial localization of recently identified mitochondrial proteins. This method can be expanded to confirm sub-mitochondrial localization and membrane topologies of previously identified mitochondrial proteins. 2017-07-07T18:14:28Z 2017-07-07T18:14:28Z 2016-05 2016-03 Article http://purl.org/eprint/type/JournalArticle 2211-1247 http://hdl.handle.net/1721.1/110551 Lee, Song-Yi; Kang, Myeong-Gyun; Park, Jong-Seok; Lee, Geunsik; Ting, Alice Y. and Rhee, Hyun-Woo. “APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest.” Cell Reports 15, 8 (May 2016): 1837–1847 © 2016 The Author(s) https://orcid.org/0000-0002-8277-5226 en_US http://dx.doi.org/10.1016/j.celrep.2016.04.064 Cell Reports Creative Commons Attribution 4.0 International License http://creativecommons.org/licenses/by/4.0/ application/pdf Elsevier Cell Reports |
| spellingShingle | Lee, Song-Yi Kang, Myeong-Gyun Park, Jong-Seok Lee, Geunsik Rhee, Hyun-Woo Ting, Alice Y APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest |
| title | APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest |
| title_full | APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest |
| title_fullStr | APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest |
| title_full_unstemmed | APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest |
| title_short | APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest |
| title_sort | apex fingerprinting reveals the subcellular localization of proteins of interest |
| url | http://hdl.handle.net/1721.1/110551 https://orcid.org/0000-0002-8277-5226 |
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