Entity

Time filter

Source Type

La Tronche, France

Liguori L.,HumProTher laboratory | Blesneac I.,CNRS Institute of Pharmacology and Structural Biology | Madern D.,CNRS Institute of Pharmacology and Structural Biology | Vivaudou M.,CNRS Institute of Pharmacology and Structural Biology | Lenormand J.-L.,HumProTher laboratory
Protein Expression and Purification | Year: 2010

The pea chloroplastic outer envelope protein OEP24 is a voltage-dependent channel that can function as a general solute channel in plants. OEP24 is a close functional homologue of VDAC which, in mammalian cells, modulates the permeability of the outer mitochondrial membrane. Here, we describe the production in a one-step reaction of active OEP24 in proteoliposomes or in soluble form using a cell-free expression system. We combine evidence from electrophysiological experiments, biophysical characterization, and biochemical analysis demonstrating that OEP24 is present as a functional channel in liposomes. Thus, production of OEP-containing proteoliposomes may provide a helpful tool for deciphering the role of the OEP family members. © 2009 Elsevier Inc. All rights reserved. Source


Varnier A.,CEA Grenoble | Kermarrec F.,CEA Grenoble | Blesneac I.,CEA Grenoble | Moreau C.,CEA Grenoble | And 3 more authors.
Journal of Membrane Biology | Year: 2010

A simple method for the reconstitution of membrane protein from submicron proteoliposomes into giant unilamellar vesicles (GUVs) is presented here: This method does not require detergents, fusion peptides or a dehydration step of the membrane protein solution. In a first step, GUVs of lipids were formed by electroformation, purified and concentrated; and in a second step, the concentrated GUV solution was added to a small volume of vesicles or proteoliposomes. Material transfer from submicron vesicles and proteoliposomes to GUVs occurred spontaneously and was characterized with fluorescent microscopy and patch-clamp recordings. As a functional test, the voltage-dependent, anion-selective channel protein was reconstituted into GUVs, and its electrophysiological activity was monitored with the patch clamp. This method is versatile since it is independent of the presence of the protein, as demonstrated by the fusion of fluorescently labeled submicron vesicles and proteoliposomes with GUVs. © 2010 Springer Science+Business Media, LLC. Source

Discover hidden collaborations