Mapping the faces of mitochondria and ER
APEX2 proximity biotinylation yields high-quality proteomic maps of cytosol-facing outer mitochondrial and ER membranes in living human cells.
Source
Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation
Study at a glance
- Design
- Animal / in-vitro — APEX2 proximity labeling of cytosol-facing OMM and ER membranes in living HEK 293T cells
- N
- SILAC proteomic maps with two replicates per membrane — cell-resource study
- Population
- HEK 293T cells expressing OMM/ERM APEX2 fusions
- Outcome
- Proteomes of cytosol-facing outer mitochondrial and ER membranes
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Mapping the faces of mitochondria and ER
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Maps enrich 22 more classes in the OMM proteome and 72 in the ERM proteome.
Our APEX2-derived proteomic maps () contain most of these protein classes, and enrich several more (22 in the OMM proteome (), and 72 in the ERM proteome ()) that were not previously known to reside at these membranes.
What method produced the membrane maps?
APEX2-derived proteomic maps () contain most of these protein classes, and enrich several more (22 in the OMM proteome (), and 72 in the ERM proteome ()) that were not previously known to reside at these membranes.
What they did
Used APEX2 proximity labeling in living cells to define proteins on the OMM and ER membrane facing the cytosol.
What they found
Maps capture expected functional classes and enrich many additional proteins (22 OMM; 72 ERM), providing resources beyond classical fractionation.
The limits
What it doesn't show
A proteomic inventory is not a full functional validation of every newly enriched protein.
Key terms
- APEX2
- Engineered peroxidase for proximity biotinylation proteomics.
- OMM
- Outer mitochondrial membrane.
- ERM
- Endoplasmic reticulum membrane.
- Proximity biotinylation
- Labeling proteins near an enzyme tag in living cells.
- Sub-organellar proteome
- Protein inventory of a membrane/compartment face.
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Method:
Common questions
Method?
APEX2 proximity biotinylation.
Membranes mapped?
Cytosol-facing OMM and ERM.
Extra enrichments reported?
22 OMM and 72 ERM classes/proteins noted.
Prior approach?
Biochemical fractionation.
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