Extracellular Vesicles: A Platform for the Structure Determination of Membrane Proteins by Cryo-EM

Tzviya Zeev-Ben-Mordehai, Daven Vasishtan, C. Alistair Siebert, Cathy Whittle, Kay Gruenewald

Research output: Contribution to journalArticleAcademicpeer-review


Membrane protein-enriched extracellular vesicles (MPEEVs) provide a platform for studying intact membrane proteins natively anchored with the correct topology in genuine biological membranes. This approach circumvents the need to conduct tedious detergent screens for solubilization, purification, and reconstitution required in classical membrane protein studies. We have applied this method to three integral type I membrane proteins, namely the Caenorhabditis elegans cell-cell fusion proteins AFF-1 and EFF-1 and the glycoprotein B (gB) from Herpes simplex virus type 1 (HSV1). Electron cryotomography followed by subvolume averaging allowed the 3D reconstruction of EFF-1 and HSV1 gB in the membrane as well as an analysis of the spatial distribution and interprotein interactions on the membrane. MPEEVs have many applications beyond structural/functional investigations, such as facilitating the raising of antibodies, for protein-protein interaction assays or for diagnostics use, as biomarkers, and possibly therapeutics.
Original languageEnglish
Pages (from-to)1687-1692
Number of pages6
JournalStructure with Folding & design
Issue number11
Publication statusPublished - 4 Nov 2014
Externally publishedYes


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