Interactions of Bax and tBid with lipid monolayers.
The release of cytochrome c from mitochondria to the cytosol is a crucial step of apoptosis that involves interactions of Bax and tBid proteins with the mitochondrial membrane. We investigated Bax and tBid interactions with (i) phosphatidylcholine (PC) monolayer as the main component of the outer leaflet of the outer membrane, (ii) with phosphatidylethanolamine (PE) and phosphatidylserine (PS) that are present in the inner leaflet and (iii) with a mixed PC/PE/Cardiolipin (CL) monolayer of the contact sites between the outer and inner membranes. These interactions were studied by measuring the increase of the lipidic monolayer surface pressure induced by the proteins. Our measurements suggest that tBid interacts strongly with the POPC/DOPE/CL, whereas Bax interaction with this monolayer is about 12 times weaker. Both tBid and Bax interact moderately half as strongly with negatively charged DOPS and non-lamellar DOPE monolayers. TBid also slightly interacts with DOPC. Our results suggest that tBid but not Bax interacts with the PC-containing outer membrane. Subsequent insertion of these proteins may occur at the PC/PE/CL sites of contact between the outer and inner membranes. It was also shown that Bax and tBid being mixed in solution inhibit their insertion into POPC/DOPE/CL monolayer. The known 3-D structures of Bax and Bid allowed us to propose a structural interpretation of these experimental results.
Van Mau
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Centre de Recherche de Biochimie Macromoléculaire CRBM, CNRS, 1919, Route de Mende, 34293 Montpellier, cedex 5, France.
Kajava
A V
AV
Bonfils
C
C
Martinou
J-C
JC
Harricane
M-C
MC
D000818:Animals; D051023:BH3 Interacting Domain Death Agonist Protein / Q000737:chemistry / Q000378:metabolism*; D002118:Calcium / Q000502:physiology; D002308:Cardiolipins / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D006801:Humans; D050356:Lipid Metabolism; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051028:bcl-2-Associated X Protein / Q000737:chemistry / Q000378:metabolism*
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; C494826:BAX protein, human; D051023:BH3 Interacting Domain Death Agonist Protein; D002308:Cardiolipins; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D051028:bcl-2-Associated X Protein; C094877:dioleoyl phosphatidylethanolamine; D002118:Calcium; C028694:1-palmitoyl-2-oleoylphosphatidylcholine