Generation of proteoliposomes from subcellular fractions

Article Article scientifique Article Generation of proteoliposomes from subcellular fractions eng 1997 author Rojo Ortiz De Lanzagorta Manuel 91130 author Budin Nicolas Philippe 83297 author Kellner Roland author Gruenberg Jean 29661 2620-2628 10.1002/elps.1150181418 9527492 0173-0835 https://onlinelibrary.wiley.com/doi/10.1002/elps.1150181418 Intracellular membranes are highly dynamic, yet they retain their identity and functional characteristics. Integral membrane proteins, which must confer this specific membrane identity, remain poorly characterized at the biochemical level, largely because detergent-mediated solubilization is required for purification and analysis, and several properties of integral membrane proteins can only be investigated when the molecule is properly embedded in a lipid bilayer. We present a method for the efficient reconstitution into proteoliposomes of integral membrane proteins from subcellular fractions. Integral membrane proteins were identified on high-resolution two-dimensional gels after selective extraction of soluble and peripheral membrane proteins; they accounted for 8% of the number of resolved polypeptides. A reconstitution procedure based on membrane solubilization with dodecyl-octaoxyethylene (C<sub>12</sub>E<sub>8</sub>) and subsequent detergent removal with BioBeads SM-2 resulted in the efficient reconstitution of several membrane proteins into proteoliposomes of uniform density. The generated proteoliposomes strongly resemble the starting membrane fraction in protein composition. This reconstitution allows the functional characterization of integral membrane proteins after enrichment and/or specific (immuno)depletion. Membrane solubilization Membrane reconstitution Proteoliposome Detergent Integral membrane protein Two-dimensional polyacrylamide gel electrophoresis Endosome Golgi Dewey 540 39 Département de biochimie 144 false Electrophoresis 18 14 ca279bd6-1d45-48a5-be1f-a261bd5b202f Article (Published version) Restricted Rojo_Electrophoresis_1997182620.pdf 7456958 application/pdf restrictedAccess Faculté des sciences / Section de chimie et biochimie / Département de biochimie

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Generation of proteoliposomes from subcellular fractions	Electrophoresis			1997	158	0

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Budin, Nicolas Philippe