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Inside‐out plant plasma membrane vesicles of high purity obtained by aqueous two‐phase partitioning
Author(s) -
Larsson Christer,
Widell Susanne,
Sommarin Marianne
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)81142-6
Subject(s) - vesicle , acridine orange , chemistry , atpase , membrane , aqueous solution , chromatography , aqueous two phase system , partition coefficient , biophysics , analytical chemistry (journal) , biochemistry , organic chemistry , biology , enzyme , apoptosis
Highly purified plasma membranes obtained from leaves of sugar beet ( Beta vulgaris L.) by aqueous two‐phase partitioning were separated into two fractions by further phase partition steps. The vesicles partitioning to the interface showed an ATP‐dependent H + ‐uptake (measured using the pH probe acridine orange) and a negligible K + ,Mg 2+ ‐ATPase latency, while the vesicles partitioning in the upper phase showed only slow H + ‐uptake and a high ATPase latency on addition of Triton X‐100. Based on these results the material at the interface is estimated to contain ∼90% sealed, inside‐out vesicles, and the material in the upper phase ∼90% sealed, right‐side‐out vesicles