Single-step production of functional OEP24 proteoliposomes.
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.
Liguori
Lavinia
L
TheREx, TIMC-IMAG Laboratory, University Joseph Fourier, UFR de Médecine, 38706 La Tronche, Cedex, France. lliguori@ujf-grenoble.fr
Blesneac
Iulia
I
Madern
Dominique
D
Vivaudou
Michel
M
Lenormand
Jean-Luc
JL
D017209:Apoptosis; D001681:Biological Assay; D015415:Biomarkers / Q000378:metabolism; D001709:Biotechnology / Q000379:methods*; D053453:Caspase 9 / Q000378:metabolism; D002470:Cell Survival; D002942:Circular Dichroism; D055724:Electrophysiological Phenomena; D004789:Enzyme Activation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D045325:HCT116 Cells; D006801:Humans; D007473:Ion Channels / Q000096:biosynthesis*; D010940:Plant Proteins / Q000096:biosynthesis*; D011510:Proteolipids / Q000378:metabolism*; D012995:Solubility
D015415:Biomarkers; D007473:Ion Channels; C113819:OEP24 protein, Pisum sativum; D010940:Plant Proteins; D011510:Proteolipids; C020485:proteoliposomes; D049452:Green Fluorescent Proteins; D053453:Caspase 9