Liposomes-mediated delivery of pro-apoptotic therapeutic membrane proteins.
The delivery of functional therapeutic proteins by lipid vesicles into targeted living cells is one of the most promising strategies for treatment of different diseases and cancer. The use of this system in the delivery of membrane proteins directly into cells remains to be tested because the methods for producing membrane proteins are difficult to perform. Here we describe the effect of proteoliposomes containing the voltage-dependent anion channel (VDAC) and pro-apoptotic Bak, both produced with an optimized cell-free expression system. For the first time, recombinant VDAC and Bak proteins are synthesized and directly integrated into the lipidic bilayer of natural liposomes in a one-step reaction. VDAC has been shown to play an essential role in apoptosis in mammalian cells by regulating cytochrome c release from mitochondria and Bak modulates mitochondrial membrane permeability upon activation. Internalization of recombinant proteoliposomes into mammalian cells induces apoptosis by release of cytochrome c and caspases activation. These results highlight that membrane proteins integrated in natural liposomes can represent an excellent candidate for cancer protein therapy.
Liguori
Lavinia
L
HumProTher laboratory, GREPI, TIMC IMAG, UMR5525, UJF/CNRS, University of Joseph Fourier, CHU-Grenoble, BP217, 38043 Grenoble Cedex 9, France.
Marques
Bruno
B
Villegas-Mendez
Ana
A
Rothe
Romy
R
Lenormand
Jean-Luc
JL
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism; D016503:Drug Delivery Systems / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D008051:Lipid Bilayers / Q000737:chemistry; D008081:Liposomes; D008565:Membrane Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D011510:Proteolipids / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D050994:Voltage-Dependent Anion Channels / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics
D008051:Lipid Bilayers; D008081:Liposomes; D008565:Membrane Proteins; D011510:Proteolipids; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53; D050994:Voltage-Dependent Anion Channels; D050998:bcl-2 Homologous Antagonist-Killer Protein; C020485:proteoliposomes; D045304:Cytochromes c; D053148:Caspase 3