Directed evolution of APEX2 for electron microscopy and proximity labeling
APEX is an engineered peroxidase that functions as an electron microscopy tag and a promiscuous labeling enzyme for live-cell proteomics. Because limited sensitivity precludes applications requiring low APEX expression, we used yeast-display evolution to improve its catalytic efficiency. APEX2 is fa...
| Main Authors: | , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
Nature Publishing Group
2017
|
| Online Access: | http://hdl.handle.net/1721.1/110613 https://orcid.org/0000-0002-2687-3470 https://orcid.org/0000-0002-8277-5226 |
| _version_ | 1826188718066630656 |
|---|---|
| author | Kamer, Kimberli J Deerinck, Thomas J Ellisman, Mark H Mootha, Vamsi K Lam, Stephanie Shih-Min Martell, Jeffrey Daniel Ting, Alice Y |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Kamer, Kimberli J Deerinck, Thomas J Ellisman, Mark H Mootha, Vamsi K Lam, Stephanie Shih-Min Martell, Jeffrey Daniel Ting, Alice Y |
| author_sort | Kamer, Kimberli J |
| collection | MIT |
| description | APEX is an engineered peroxidase that functions as an electron microscopy tag and a promiscuous labeling enzyme for live-cell proteomics. Because limited sensitivity precludes applications requiring low APEX expression, we used yeast-display evolution to improve its catalytic efficiency. APEX2 is far more active in cells, enabling the use of electron microscopy to resolve the submitochondrial localization of calcium uptake regulatory protein MICU1. APEX2 also permits superior enrichment of endogenous mitochondrial and endoplasmic reticulum membrane proteins. |
| first_indexed | 2024-09-23T08:03:57Z |
| format | Article |
| id | mit-1721.1/110613 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:03:57Z |
| publishDate | 2017 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/1106132022-09-23T10:39:31Z Directed evolution of APEX2 for electron microscopy and proximity labeling Kamer, Kimberli J Deerinck, Thomas J Ellisman, Mark H Mootha, Vamsi K Lam, Stephanie Shih-Min Martell, Jeffrey Daniel Ting, Alice Y Massachusetts Institute of Technology. Department of Chemistry Lam, Stephanie Shih-Min Martell, Jeffrey Daniel Ting, Alice Y APEX is an engineered peroxidase that functions as an electron microscopy tag and a promiscuous labeling enzyme for live-cell proteomics. Because limited sensitivity precludes applications requiring low APEX expression, we used yeast-display evolution to improve its catalytic efficiency. APEX2 is far more active in cells, enabling the use of electron microscopy to resolve the submitochondrial localization of calcium uptake regulatory protein MICU1. APEX2 also permits superior enrichment of endogenous mitochondrial and endoplasmic reticulum membrane proteins. National Institutes of Health (U.S.) (DP1 OD003961) 2017-07-11T13:04:21Z 2017-07-11T13:04:21Z 2014-11 2014-03 Article http://purl.org/eprint/type/JournalArticle 1548-7091 1548-7105 http://hdl.handle.net/1721.1/110613 Lam, Stephanie S; Martell, Jeffrey D; Kamer, Kimberli J et al. "Directed evolution of APEX2 for electron microscopy and proximity labeling." Nature Methods 12, 1: 51–54 (January 2015) © 2015 Nature America, Inc https://orcid.org/0000-0002-2687-3470 https://orcid.org/0000-0002-8277-5226 en_US http://dx.doi.org/10.1038/nmeth.3179 Nature Methods Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Nature Publishing Group PMC |
| spellingShingle | Kamer, Kimberli J Deerinck, Thomas J Ellisman, Mark H Mootha, Vamsi K Lam, Stephanie Shih-Min Martell, Jeffrey Daniel Ting, Alice Y Directed evolution of APEX2 for electron microscopy and proximity labeling |
| title | Directed evolution of APEX2 for electron microscopy and proximity labeling |
| title_full | Directed evolution of APEX2 for electron microscopy and proximity labeling |
| title_fullStr | Directed evolution of APEX2 for electron microscopy and proximity labeling |
| title_full_unstemmed | Directed evolution of APEX2 for electron microscopy and proximity labeling |
| title_short | Directed evolution of APEX2 for electron microscopy and proximity labeling |
| title_sort | directed evolution of apex2 for electron microscopy and proximity labeling |
| url | http://hdl.handle.net/1721.1/110613 https://orcid.org/0000-0002-2687-3470 https://orcid.org/0000-0002-8277-5226 |
| work_keys_str_mv | AT kamerkimberlij directedevolutionofapex2forelectronmicroscopyandproximitylabeling AT deerinckthomasj directedevolutionofapex2forelectronmicroscopyandproximitylabeling AT ellismanmarkh directedevolutionofapex2forelectronmicroscopyandproximitylabeling AT moothavamsik directedevolutionofapex2forelectronmicroscopyandproximitylabeling AT lamstephanieshihmin directedevolutionofapex2forelectronmicroscopyandproximitylabeling AT martelljeffreydaniel directedevolutionofapex2forelectronmicroscopyandproximitylabeling AT tingalicey directedevolutionofapex2forelectronmicroscopyandproximitylabeling |