Molecular species of cardiolipin in relation to other mitochondrial phospholipids. Is there an acyl specificity of the interaction between cardiolipin and the ADP/ATP carrier?
Molecular species in the three major mitochondrial lipids cardiolipin, phosphatidylcholine and phosphatidylethanolamine were analysed in bovine heart and Saccharomyces cerevisiae. In both organisms cardiolipin contains mainly diacylglycerol moieties with two unsaturated chains and a significant higher proportion of C18-C18 species than phosphatidylcholine and phosphatidylethanolamine. To study whether the specific acyl composition of cardiolipin has a functional significance in lipid-protein interaction, experiments were made with the isolated ADP/ATP carrier of bovine heart mitochondria since this dimeric protein is known to be tightly associated with six molecules of cardiolipin [Beyer, K. and Klingenberg, M. (1985) Biochemistry 24, 3821-3826]. This association seems to be very strong as protein-bound cardiolipin does not exchange with soluble cardiolipin on a time scale of hours. Analysis of the species composition suggests that one carriers dimer is associated with four molecules of tetralinoleoyl cardiolipin and two molecules of trilinoleoyl-monolinolenoyl cardiolipin. Catalytic hydrogenation of the acyl chains of carrier-bound cardiolipin does not result in release of cardiolipin as judged by 31P-NMR spectroscopy. The ADP/ATP carrier was reconstituted with saturated phosphatidylcholines and spin-labelled cardiolipin whose double bonds were subsequently saturated by catalytic hydrogenation. ESR spectroscopy shows that saturation of spin-labelled cardiolipin has no significant impact on its association with the ADP/ATP carrier. However, precipitation of the detergent-solubilized ADP/ATP carrier can only be induced by addition of unsaturated but not by saturated cardiolipin. It is concluded that the specific acyl composition of cardiolipin is not a prerequisite of its high affinity for the ADP/ATP carrier, at least when the protein is reconstituted in a saturated phosphatidylcholine environment.
Schlame
M
M
Institute of Physical Biochemistry, University of Munich, Federal Republic of Germany.
Beyer
K
K
Hayer-Hartl
M
M
Klingenberg
M
M
D000818:Animals; D002308:Cardiolipins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D002417:Cattle; D004578:Electron Spin Resonance Spectroscopy; D005227:Fatty Acids / Q000032:analysis; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008928:Mitochondria / Q000737:chemistry* / Q000201:enzymology; D008929:Mitochondria, Heart / Q000737:chemistry* / Q000201:enzymology; D000226:Mitochondrial ADP, ATP Translocases / Q000378:metabolism*; D010743:Phospholipids / Q000494:pharmacology; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000032:analysis / Q000201:enzymology; D013113:Spin Labels; D013329:Structure-Activity Relationship
D002308:Cardiolipins; D005227:Fatty Acids; D010743:Phospholipids; D013113:Spin Labels; D000226:Mitochondrial ADP, ATP Translocases