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CryoAPEX - an electron tomography tool for subcellular localization of membrane proteins.

J Cell Sci. 2019-03; 
SenguptaRanjan,PoderyckiMichael J,MattooS
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Molecular Biology Reagents The HYPE–APEX2 plasmid was ordered through GenScript. Get A Quote

摘要

We describe a method, termed cryoAPEX, which couples chemical fixation and high-pressure freezing of cells with peroxidase tagging (APEX) to allow precise localization of membrane proteins in the context of a well-preserved subcellular membrane architecture. Further, cryoAPEX is compatible with electron tomography. As an example, we apply cryoAPEX to obtain a high-resolution three-dimensional contextual map of the human FIC (filamentation induced by cAMP) protein, HYPE (also known as FICD). HYPE is a single-pass membrane protein that localizes to the endoplasmic reticulum (ER) lumen and regulates the unfolded protein response. Alternate cellular locations for HYPE have been suggested. CryoAPEX analysi... More

关键词

CryoAPEX,Cryofixation,Electron tomography,FIC AMPylation,Membrane pro