The oxidized phospholipid PazePC modulates interactions between Bax and mitochondrial membranes.
Activation of the pro-apoptotic protein Bax under intracellular oxidative stress is closely related to its association with the mitochondrial outer membrane (MOM) system, ultimately resulting in cell death. The precise mechanism by which this activation and the subsequent structural changes in the protein occur is currently unknown. In addition to triggering the onset of apoptosis, oxidative stress generates oxidized lipids whose impact on mitochondrial membrane integrity and the activity of membrane-associated Bax is unclear. We therefore devised a model system that mimics oxidative stress conditions by incorporating oxidized phospholipids (OxPls) into mitochondria-like liposomes, and studied the OxPls' impact on Bax-membrane interactions. Differential scanning calorimetry (DSC) was used to study membrane organization and protein stability, while conformational changes in the protein upon contact with lipid vesicles were monitored using far-UV circular dichroism (CD) spectroscopy. The thermograms for liposomes containing the OxPl 1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine (PazePC) differed dramatically from those for unmodified liposomes. Moreover, Bax exhibited enhanced thermal stability in the presence of the modified liposomes, indicating that it interacted strongly with PazePC-containing membranes. The presence of PazePC also increased the α-helical character of Bax compared to the protein alone or with PazePC-free vesicles, at 10°C, 20°C, and 37°C. Presumably, the presence of PazePC-like OxPls a) increases the population of membrane-associated Bax and b) facilitates the protein's insertion into the membrane by distorting the bilayer's organization, as seen by solid-state high-resolution (1)H and (31)P magic angle spinning nuclear magnetic resonance (MAS NMR) spectroscopy.
Department of Chemistry, University of Umeå, Umeå, Sweden.
Marcus
M
Wallgren
Martin
M
Lidman
Quoc Dat
QD
Pham
Konrad
K
Cyprych
Gerhard
G
Gröbner
D017209:Apoptosis; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008928:Mitochondria / Q000378:metabolism*; D018384:Oxidative Stress; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism*
C528536:1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine; D011994:Recombinant Proteins; D051028:bcl-2-Associated X Protein; D010767:Phosphorylcholine