Journal
NATURE METHODS
Volume 12, Issue 1, Pages 51-54Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nmeth.3179
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Funding
- US National Institutes of Health [Dp1 0D003961, P41 GM103412, R01GM086197, 5R01GM077465-08]
- Howard Hughes Medical Institute
- Howard Hughes Medical Institute Collaborative Initiative Award
- US National Science Foundation
- National Defense Science and Engineering Graduate Fellowships
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM086197, R01GM077465, P41GM103412] Funding Source: NIH RePORTER
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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.
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