Cardiolipins

Cardiolipin (IUPAC name 1,3-bis(sn-3’-phosphatidyl)-sn-glycerol, also known as Calcutta antigen) is an important component of the inner mitochondrial membrane, where it constitutes about 20% of the total lipid composition. It can also be found in the membranes of most bacteria. The name 'cardiolipin' is derived from the fact that it was first found in animal hearts. It was first isolated from beef heart in the early 1940s. In mammalian cells, but also in plant cells, cardiolipin (CL) is found almost exclusively in the inner mitochondrial membrane where it is essential for the optimal function of numerous enzymes that are involved in mitochondrial energy metabolism.Cardiolipin bicyclic structureStructure of NAONAO & CL arranged in a highly ordered way Cardiolipin (IUPAC name 1,3-bis(sn-3’-phosphatidyl)-sn-glycerol, also known as Calcutta antigen) is an important component of the inner mitochondrial membrane, where it constitutes about 20% of the total lipid composition. It can also be found in the membranes of most bacteria. The name 'cardiolipin' is derived from the fact that it was first found in animal hearts. It was first isolated from beef heart in the early 1940s. In mammalian cells, but also in plant cells, cardiolipin (CL) is found almost exclusively in the inner mitochondrial membrane where it is essential for the optimal function of numerous enzymes that are involved in mitochondrial energy metabolism. Cardiolipin (CL) is a kind of diphosphatidylglycerol lipid. Two phosphatidic acid moieties connect with a glycerol backbone in the center to form a dimeric structure. So it has four alkyl groups and potentially carries two negative charges. As there are four distinct alkyl chains in cardiolipin, the potential for complexity of this molecule species is enormous. However, in most animal tissues, cardiolipin contains 18-carbon fatty alkyl chains with 2 unsaturated bonds on each of them. It has been proposed that the (18:2)4 acyl chain configuration is an important structural requirement for the high affinity of CL to inner membrane proteins in mammalian mitochondria. However, studies with isolated enzyme preparations indicate that its importance may vary depending on the protein examined. Since there are two phosphates in the molecule, each of them can catch one proton. Although it has a symmetric structure, ionizing one phosphate happens at a very different levels of acidity than ionizing both: pK1 = 3 and pK2 > 7.5. So under normal physiological conditions (wherein pH is around 7), the molecule may carry only one negative charge. The hydroxyl groups (–OH and –O−) on phosphate would form a stable intramolecular hydrogen bond with the centered glycerol's hydroxyl group, thus forming a bicyclic resonance structure. This structure traps one proton, which is quite helpful for oxidative phosphorylation. As the head group forms such compact bicycle structure, the head group area is quite small relative to the big tail region consisting of 4 acyl chains. Based on this special structure, the fluorescent mitochondrial indicator, nonyl acridine orange (NAO) was introduced in 1982, and was later found to target mitochondria by binding to CL. NAO has a very large head and small tail structure which can compensate with cardiolipin's small head large tail structure, and arrange in a highly ordered way. Several studies were published utilizing NAO both as a quantitative mitochondrial indicator and an indicator of CL content in mitochondria. However, NAO is influenced by membrane potential and/or the spatial arrangement of CL, so it's not proper to use NAO for CL or mitochondria quantitative studies of intact respiring mitochondria. But NAO still represents a simple method of assessing CL content. In eukaryotes such as yeasts, plants and animals, the synthesis processes are believed to happen in mitochondria. The first step is the acylation of glycerol-3-phosphate by a glycerol-3-phosphate acyltransferase. Then acylglycerol-3-phosphate can be once more acylated to form a phosphatidic acid (PA). With the help of the enzyme CDP-DAG synthase (CDS) (phosphatidate cytidylyltransferase), PA is converted into cytidinediphosphate-diacylglycerol (CDP-DAG). The following step is conversion of CDP-DAG to phosphatidylglycerol phosphate (PGP) by the enzyme PGP synthase, followed by dephosphorylation by PTPMT1 to form PG. Finally, a molecule of CDP-DAG is bound to PG to form one molecule of cardiolipin, catalyzed by the mitochondria-localized enzyme cardiolipin synthase (CLS). In prokaryotes such as bacteria, diphosphatidylglycerol synthase catalyses a transfer of the phosphatidyl moiety of one phosphatidylglycerol to the free 3'-hydroxyl group of another, with the elimination of one molecule of glycerol, via the action of an enzyme related to phospholipase D. The enzyme can operate in reverse under some physiological conditions to remove cardiolipin. Catabolism of cardiolipin may happen by the catalysis of phospholipase A2 (PLA) to remove fatty acyl groups. Phospholipase D (PLD) in the mitochondrion hydrolyses cardiolipin to phosphatidic acid. Because of cardiolipin's unique structure, a change in pH and the presence of divalent cations can induce a structural change. CL shows a great variety of forms of aggregates. It is found that in the presence of Ca2+ or other divalent cations, CL can be induced to have a lamellar-to-hexagonal (La-HII) phase transition. And it is believed to have a close connection with membrane fusion. The enzyme cytochrome c oxidase or Complex IV is a large transmembrane protein complex found in bacteria and the mitochondrion. It is the last enzyme in the respiratory electron transport chain of mitochondria (or bacteria) located in the mitochondrial (or bacterial) membrane. It receives an electron from each of four cytochrome c molecules, and transfers them to one oxygen molecule, converting molecular oxygen to two molecules of water. Complex IV has been shown to require two associated CL molecules in order to maintain its full enzymatic function. Cytochrome bc1(Complex III) also needs cardiolipin to maintain its quaternary structure and functional role. Complex V of the oxidative phosphorylation machinery also displays high binding affinity for CL, binding four molecules of CL per molecule of complex V.