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Cell‐Derived Vesicles for Single‐Molecule Imaging of Membrane Proteins
Author(s) -
Moonschi Faruk H.,
Effinger Amy K.,
Zhang Xiaolu,
Martin William E.,
Fox Ashley M.,
Heidary David K.,
DeRouchey Jason E.,
Richards Christopher I.
Publication year - 2015
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201408707
Subject(s) - vesicle , receptor , biophysics , chemistry , membrane , force spectroscopy , fluorescence correlation spectroscopy , cell membrane , molecule , cell surface receptor , biochemistry , biology , organic chemistry
A new approach is presented for the application of single‐molecule imaging to membrane receptors through the use of vesicles derived from cells expressing fluorescently labeled receptors. During the isolation of vesicles, receptors remain embedded in the membrane of the resultant vesicles, thus allowing these vesicles to serve as nanocontainers for single‐molecule measurements. Cell‐derived vesicles maintain the structural integrity of transmembrane receptors by keeping them in their physiological membrane. It was demonstrated that receptors isolated in these vesicles can be studied with solution‐based fluorescence correlation spectroscopy (FCS) and can be isolated on a solid substrate for single‐molecule studies. This technique was applied to determine the stoichiometry of α3β4 nicotinic receptors. The method provides the capability to extend single‐molecule studies to previously inaccessible classes of receptors.