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Abstract
A procedure for the production of plasma membrane vesicles, consisting of enzyme-assisted physical homogenization, differential centrifugation and partitioning in an aqueous polymer two phase system resulted in a fraction containing highly purified plasma membranes, as determined by vanadate-inhibited H + ATPase and near absence of intracellular membrane marker activities. H + ATPase latency tests indicated that at least 90% of the vesicles were oriented with the apoplastic surface facing outward (right-side-out orientation) in the upper phase, while inside out vesicles and other microsomal membrane material collected in the lower phase.
Published Version
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