PMID: 9432006Feb 7, 1998Paper

Detergent-mediated reconstitution of a glycosyl-phosphatidylinositol-protein into liposomes

European Journal of Biochemistry
M AngrandB Roux

Abstract

A three-step detergent-mediated reconstitution has been applied to the incorporation of a glycosyl-phosphatidylinositol-protein into liposomes. The protein studied was alkaline phosphatase from bovine intestine. Liposomes prepared by dialysis were treated with various amounts of two detergents, either n-octyl beta-D-glucoside or Triton X-100. At different steps of the solubilization process, protein was added and the detergent was removed by hydrophobic resins. The most efficient reconstitutions were obtained with an octyl glucoside concentration corresponding to the onset of liposome solubilization and with a Triton X-100 concentration leading to partial solubilization of the liposomes. The involvement of the glycosyl-phosphatidylinositol anchor in alkaline phosphatase reconstitution was demonstrated by the inability of phosphoinositol-specific phospholipase-C-hydrolysed alkaline phosphatase to incorporate into liposomes. Between 70-85% of the protein associated with liposomes were anchored in the outer leaflet of the bilayer, oriented towards the outside of the liposome. The remainder was trapped within the lumen of the liposomes.

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Citations

Aug 1, 2007·European Biophysics Journal : EBJ·Sylvain TrépoutOlivier Lambert
May 16, 2002·The International Journal of Biochemistry & Cell Biology·Fernando L CamoleziPietro Ciancaglini
May 3, 2002·European Journal of Biochemistry·Raghda LahdoLaurence de La Fournière-Bessueille
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Jul 7, 2020·Mechanisms of Ageing and Development·Günter A MüllerTimo D Müller
Jul 31, 2007·Langmuir : the ACS Journal of Surfaces and Colloids·Marie-Cécile GiocondiChristian Le Grimellec
Apr 12, 2002·Biochemical and Biophysical Research Communications·Muriel BortolatoBernard Roux
Apr 18, 2002·Biochimica Et Biophysica Acta·Frédéric RonzonBernard Roux
Aug 15, 2002·Biochimica Et Biophysica Acta·Sandrine MorandatBernard Roux
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Jan 4, 2013·Langmuir : the ACS Journal of Surfaces and Colloids·Y KaufmanV Freger
May 18, 2005·Langmuir : the ACS Journal of Surfaces and Colloids·Benjamin CrossJean-Paul Rieu

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