title: Production of plasma membrane vesicles with chloride salts and their utility as a cell membrane mimetic for biophysical characterization of membrane protein interactions.
abstract: Plasma membrane derived vesicles are used as a model system for the biochemical and biophysical investigations of membrane proteins and membrane organization. The most widely used vesiculation procedure relies on formaldehyde and dithiothreitol (DTT), but these active chemicals may introduce artifacts in the experimental results. Here we describe a procedure to vesiculate Chinese hamster ovary (CHO) cells, widely used for the expression of recombinant proteins, using a hypertonic vesiculation buffer containing chloride salts and no formaldehyde or DTT. We characterize the size distribution of the produced vesicles. We also show that these vesicles can be used for the biophysical characterization of interactions between membrane proteins.
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, Maryland 21218, United States.
authors.forename: Nuala
authors.initials: N
authors.lastname: Del Piccolo
authors.forename: Jesse
authors.initials: J
authors.lastname: Placone
authors.forename: Lijuan
authors.initials: L
authors.lastname: He
authors.forename: Sandra Carolina
authors.initials: SC
authors.lastname: Agudelo
authors.forename: Kalina
authors.initials: K
authors.lastname: Hristova
mesh_terms: D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D002712:Chlorides / Q000378:metabolism*; D006224:Cricetinae; D008565:Membrane Proteins / Q000378:metabolism*; D025941:Protein Interaction Mapping
chemical_list: D002712:Chlorides; D008565:Membrane Proteins