Does anyone know how to prepare inside-out membrane vesicles?

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Biplab Bose

A method is described in Methods in Enzymology Vol.:31 (1974) 172-180

M. R. Mozafari

Hi,

Your question remind me of "inverted micelles" or "reverse micelles":

Reverse micelles are particles dispersed in organic media obtained by the action of surfactants. Surfactant molecules organize with the polar part to the inner side able to solubilize water and the apolar part in contact with the organic solvent.

Also, you may find these papers useful:

Processing by inverted plasma membrane vesicles of in vitro ... - NCBI

https://www.ncbi.nlm.nih.gov/pubmed/3282924

by SS Krishnabhakdi - ‎1988 - ‎Cited by 6 - ‎Related articles

FEBS Lett. 1988 Apr 11;231(1):99-101. Processing by inverted plasma membrane vesicles of in vitro synthesized major lipoprotein from Escherichia coli.

Active transport of calcium in inverted membrane vesicles of ... - NCBI

https://www.ncbi.nlm.nih.gov/pubmed/4373740

by BP Rosen - ‎1974 - ‎Cited by 105 - ‎Related articles

Proc Natl Acad Sci U S A. 1974 Dec;71(12):5042-6. Active transport of calcium in inverted membrane vesicles of Escherichia coli. Rosen BP, McClees JS.

.....

Characterization of energetically functional inverted membrane ...

www.sciencedirect.com/science/article/pii/0014579394012458

by S Schrempp - ‎1994 - ‎Cited by 2 - ‎Related articles

These membrane vesicles may be the starting point for in vitro translocation ... The central part of an in vitro system is inverted membrane vesicles derived from ...

Mikhail Bogdanov

Method for preparation of uniformly oriented and sealed inside out vesicles (ISOv) from Escherichia coli cells was further developed and published by our group in Bogdanov, M., Heacock, P. and Dowhan, W.: A polytopic membrane protein displays a reversible topology dependent on membrane lipid composition. EMBO J. 21: 2107-2116, 2002. To prepare ISO vesicles Escherichia coli cells were ruptured using a French press at 560 kg/cm2 (8000 p.s.i. THIS PRESSURE IS CRUCIAL!). Orientation of membrane vesicles was verified by new vesicle sidedness assay by taking advantage of mapping of uniform topology of essential E. coli protein leader peptidase (Lep). Please see Materials and Methods and Supplemental material to EMBO J. 21: 2107-2116, 2002 manuscript for details of both protocols respectively.