Production of plasma membrane vesicles with chloride salts and their utility as a cell membrane mimetic for biophysical characterization of membrane protein interactions

Nuala Del Piccolo, Jesse Placone, Lijuan He, Sandra Carolina Agudelo, Kalina Hristova

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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.

Original languageEnglish (US)
Pages (from-to)8650-8655
Number of pages6
JournalAnalytical chemistry
Volume84
Issue number20
DOIs
StatePublished - Oct 16 2012

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ASJC Scopus subject areas

  • Analytical Chemistry

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