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1.
Metabolism of Phosphatidylglycerol, Lysylphosphatidylglycerol, and Cardiolipin of Staphylococcus aureus 总被引:5,自引:4,他引:1
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Staphylococcus aureus accumulated cardiolipin (CL) and lost phosphatidylglycerol (PG) during the stationary phase of growth. The minor lipids, phosphatidylethanolamine and phosphatidylglucose, also accumulated, whereas the lysylphosphatidylglycerol (LPG) content of the membrane remained constant as stationary phase continued. During exponential growth, the proportions and total content of phospholipids per cell remained constant. The metabolism of the phospholipids was examined under these conditions. In pulse-chase experiments, the phospholipids lost (14)C from the glycerols slower than (32)P. When the phospholipids were labeled with (14)C glycerol, the unacylated glycerols of PG and LPG lost (14)C, whereas the diacylated glycerols either accumulated or did not lose (14)C. In all experiments, the PG showed a more rapid metabolism than the LPG. When staphylococcal CL was hydrolyzed by Haemophilus parainfluenzae CL-specific phospholipase D into phosphatidic acid (PA) and PG, the incorporation of (32)P into both of the phosphates of CL was found to be parallel at both the PG and PA ends of the molecule. However, the specific activity of the (32)P at the PA end was twice that at the PG end of the molecule. The PG end of the CL apparently came from a portion of the cellular PG pool with about 20% the specific activity of the total cellular PG. The turnover of two of the glycerols of the PG portion of CL was like that of the cellular PG. The diacylated glycerol of the PG and of CL and of the membrane PG showed neither turnover nor incorporation of (14)C. Half of the radioactivity was lost from the middle glycerol of CL and the free glycerol of the cellular PG in one bacterial doubling. The diacylated glycerol from the other end of the CL molecule (the PA end) lost radioactivity almost as rapidly as the middle glycerol for 10 min. After the initial rapid loss, the turnover slowed to a rate 10 times slower than the middle glycerol, indicating that the (14)C was actually accumulating at this end of the molecule. The phosphates and glycerols involved in the hydrolysis and resynthesis of the CL molecule during exponential growth in S. aureus apparently come from different pools of PG. 相似文献
2.
Vreken P Valianpour F Nijtmans LG Grivell LA Plecko B Wanders RJ Barth PG 《Biochemical and biophysical research communications》2000,279(2):378-382
Cardiolipin (CL) and phosphatidylglycerol (PG) are the major polyglycerophospholipids observed in mammalian tissues. CL is exclusively found in the inner mitochondrial membrane and is required for optimal function of many of the respiratory and ATP-synthesizing enzymes. The role of CL in oxidative phosphorylation is, however, not fully understood and although reduced CL content leads to aberrant cell function, no human disorders with a primary defect in cardiolipin metabolism have been described. In this paper we present evidence that patients with the rare disorder X-linked cardioskeletal myopathy and neutropenia (Barth syndrome, MIM 302060) have a primary defect in CL and PG remodeling. We investigated phospholipid metabolism in cultured skin fibroblasts of patients and show that the biosynthesis rate of PG and CL is normal but that the CL pool size is 75% reduced, indicating accelerated degradation. Moreover, the incorporation of linoleic acid, which is the characteristic acyl side chain found in mammalian CL, into both PG and CL is significantly reduced, whereas the incorporation of other fatty acids into these phospholipids is normal. We show that this defect was only observed in Barth syndrome patients' cells and not in cells obtained from patients with primary defects in the respiratory chain, demonstrating that the observed defect is not secondary to respiratory chain dysfunction. These results imply that the G4.5 gene product, which is mutated in Barth syndrome patients, is specifically involved in the remodeling of PG and CL and for the first time identify an essential factor in this important cellular process. 相似文献
3.
Phospholipid Metabolism in the Absence of Net Phospholipid Synthesis in a Glycerol-Requiring Mutant of Bacillus subtilis 总被引:8,自引:5,他引:3
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A glycerol-requiring auxotroph of Bacillus subtilis showed no net synthesis of phospholipid when deprived of glycerol. Although there was no net synthesis of phospholipid, we found that: (i) fatty acids and (32)P were slowly incorporated into phospholipid; (ii) in pulse-chase experiments, both (32)P and (14)C in the glycerol portion of the phospholipids were lost from phosphatidlyglycerol (PG) and lysylphosphatidylglycerol and accumulated in cardiolipin (CL); (iii) the proportions of the phospholipids in the membrane changed with a loss of PG and an accumulation of CL. The addition of glycerol to the glycerol-deprived cells resulted in a rapid incorporation of glycerol and restoration to the predeprivation metabolism and PG to CL ratio. 相似文献
4.
Consequences of the inhibition of cardiolipin metabolism in Haemophilus parainfluenzae 总被引:4,自引:3,他引:1
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Examination of phospholipid metabolism in Haemophilus parainfluenzae with inhibitors of various cellular functions indicated that macromolecular synthesis and lipid metabolism can be dissociated at least for a short time. Two classes of inhibitors have relatively specific effects on cardiolipin (CL) metabolism. Pentachlorophenol and p-hydroxymercuribenzoate blocked CL synthesis but allowed CL hydrolysis to phosphatidic acid and phosphatidyl glycerol (PG); 3,3',4,5'-tetrachlorosalicylanilide (TCS) and carbonyl cyanide m-chlorophenylhydrazone (m-CCCP) blocked CL hydrolysis with the stoichiometric accumulation of CL. It appeared as if TCS and m-CCCP inhibited a vital activity coupled with the hydrolysis of CL by the highly active, CL-specific phospholipase D found in this organism. Because TCS and m-CCCP are thought to act by destroying the proton gradient thereby interrupting energy-dependent transport, it is possible that a highly active portion of the cellular CL could be coupled to some phase of this process. 相似文献
5.
Hullin-Matsuda F Kawasaki K Delton-Vandenbroucke I Xu Y Nishijima M Lagarde M Schlame M Kobayashi T 《Journal of lipid research》2007,48(9):1997-2008
Bis(monoacylglycero)phosphate (BMP) is a unique lipid enriched in the late endosomes participating in the trafficking of lipids and proteins through this organelle. The de novo biosynthesis of BMP has not been clearly demonstrated. We investigated whether phosphatidylglycerol (PG) and cardiolipin (CL) could serve as precursors of de novo BMP synthesis using two different cellular models: CHO cells deficient in phosphatidylglycerophosphate (PGP) synthase, the enzyme responsible for the first step of PG synthesis; and human lymphoblasts from patients with Barth syndrome (BTHS), characterized by mutations in tafazzin, an enzyme implicated in the deacylation-reacylation cycle of CL. The biosynthesis of both PG and BMP was reduced significantly in the PGP synthase-deficient CHO mutants. Furthermore, overexpression of PGP synthase in the deficient mutants induced an increase of BMP biosynthesis. In contrast to CHO mutants, BMP biosynthesis and its fatty acid composition were not altered in BTHS lymphoblasts. Our results thus suggest that in mammalian cells, PG, but not CL, is a precursor of the de novo biosynthesis of BMP. Despite the decrease of de novo synthesis, the cellular content of BMP remained unchanged in CHO mutants, suggesting that other pathway(s) than de novo biosynthesis are also used for BMP synthesis. 相似文献
6.
Cardiolipin-specific phospholipase D of Haemophilus parainfluenzae. II. Characteristics and possible significance 总被引:5,自引:5,他引:0
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A phospholipase specific for cardiolipin (CL) was found in the membrane of Haemophilus parainfluenzae. The enzyme hydrolyzed CL to phosphatidic acid (PA) and phosphatidylglycerol (PG), indicating that it was a phospholipase D (an enzyme activity believed to be confined to higher plants). In addition to its substrate specificity, this enzyme was unusual in its requirement for Mg(2+) (K(m) of 1.3 mm) for maximal activity and its inhibition by chelating agents, heavy metals, some detergents, and organic solvents. When inhibitors of phospholipase activity were added to the growth medium, CL accumulated and PG disappeared in the membrane, suggesting that the phospholipase D was active in vivo. The activity of phospholipase D in cell-free homogenates was greater than expected from earlier studies of CL metabolism and greater than the other phospholipase activities detected in the homogenate. The high activity of the CL-specific phospholipase D suggests there might be a very active degradation of CL to PG and PA and an active resynthesis of CL from the hydrolysis products. 相似文献
7.
In Escherichia coli membranes, three major phospholipids are formed: phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and cardiolipin (CL). We report here the survival of mutants lacking either PE or both PG and CL at an acid pHo and during long-term survival experiments. Stationary phase cultures of E. coli lacking PE are much more sensitive to acid shock (pHo 3) than the wild-type strain. Moreover, in the strain lacking PE, long-term survival in stationary phase is impaired and after 5 days no viable cells are recovered. The survival of an exponential phase culture to acid shock is known to be increased if the culture is exposed to moderately acid conditions (pHo 5) prior to a shift to pHo 3. If either PE or both PG and CL are missing, the exposure to pHo 5 does not increase the survival at pHo 3. 相似文献
8.
Phospholipid metabolism during changes in the proportions of membrane-bound respiratory pigments in Haemophilus parainfluenzae 总被引:11,自引:9,他引:2
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After a transition from high to low oxygen tension, there was a twofold to 50-fold increase in the content of membrane-bound respiratory pigments of Haemophilus parainfluenzae, and there were concurrent changes in the metabolism of the membrane phospholipids: (i) a twofold decrease in the rate of turnover of the phosphate in all the phospholipids; (ii) a shift from simple one-phase, linear incorporation of phosphate into phospholipids to a complex biphasic incorporation of phosphate into phospholipids; and (iii) an increase in the total phospholipids with a slight increase in the proportion of phosphatidylglycerol (PG) and a slight decrease in the proportion of phosphatidylethanolamine (PE). Changes in the rates of incorporation of phosphate into the phospholipids occurred without a change in the rate of bacterial growth. When the compensatory adjustment of the proportions of the respiratory pigments reached a steady state, the total phospholipid, the rate of incorporation of phosphate into phospholipids, and the proportion of PG fell. At steady-state proportions of cytochromes, the proportion of PE and the rate of turnover of the phosphate in the phospholipids increased. All through an incorporation experiment of 1.5 divisions, the specific activity of the phosphate of PG was twice that of phosphatidic acid (PA). The phosphate of PG turned over 1.2 to 1.5 times more rapidly than the phosphate of PA in cells with high and low cytochrome levels. If the PA was an accurate measure of the precursor for the cytidine-5′-diphosphate-diglyceride, which in turn was the precursor of all the lipids, then the results of these experiments suggested that exchange reactions, in addition to synthesis from PA, were involved in phospholipid metabolism. These reactions were more sensitive to changes in oxygen concentration than was the growth rate. 相似文献
9.
Two distinct pools of membrane phosphatidylglycerol in Bacillus megaterium. 总被引:4,自引:4,他引:0
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The predominant membrane lipid in Bacillus megaterium ATCC 14581, phosphatidylglycerol (PG), is present in two distinct pools, as shown by [32P]phosphate incorporation and chase experiments. One pool (PGt) undergoes rapid turnover of the phosphate moiety, whereas the second pool (PGs) exhibits metabolic stability in this group. The phosphate moiety of the other major phospholipid, phosphatidylethanolamine, is stable to turnover. [32P]phosphate- and [2-3H]glycerol-equilibrated cultures yielded the following glycerolipid composition: 56 mol% PG (34 mol% PGt and 22 mol% PGs), 21 mol% phosphatidylethanolamine, 1 to 2 mol% phosphatidylserine, 20 mol% diglycerides, less than 0.5 mol% cardiolipin, and 0.2 to 0.4 mol% lysophosphatidylglycerol. Accumulation of PG was halted immediately after the addition of cerulenin, an inhibitor of de novo fatty acid synthesis, whereas phosphatidylethanolamine accumulation continued at the expense of the diglyceride and PG pools. Strikingly, initial rates of [32P]phosphate incorporation into PG were unaffected by cerulenin. In control cultures at 35 degrees C, incorporation of [32P]phosphate into PG exhibited a biphasic time course, whereas incorporation into phosphatidylethanolamine was concave upward and lagged behind that of PG during the initial rapid phase of PG incorporation. Finally, levels of lysophosphatidylglycerol expanded rapidly after cerulenin addition at 20 degrees C, but not at 35 degrees C. Moreover, incorporation of [32P]phosphate into lysophosphatidylglycerol lagged behind incorporation into PG in both the presence and absence of cerulenin at 20 and 35 degrees C. 相似文献
10.
An analysis of the turnover of the major phospholipids of Micrococcus cerificans growing or nongrowing cultures. The turnover rates of (14)C-PE and (14)C-PE were 61.5% of the total phospholipid, exhibited no significant rate of turnover in either growing or nongrowing cultures. The turnover rates of PE-(14)C and PE-(32)P were 3.2% per hr and 1.2% per hr, respectively. Phosphatidylglycerol (PG) exhibited a turnover rate of 11% and 7.7% per hr for (14)C and (32)P, respectively, indicating an extremely slow metabolism. PG metabolism was examined in greater detail, and the data indicated a preferential 75% incorporation of glycerol-1,3-(14)C into the unacylated portion of the PG molecule. The turnover of cardiolipin (CL) was extremely slow in growing cells whereas nongrowing cells exhibited a 30% and 36% increase per hr for (14)C-Cl and (14)C-CL, respectively. Glycerol-1,3-(14)C was not converted to phospholipid fatty acid carbon; all radioactivity appeared only in the water-soluble backbone of the phospholipids. The kinetics of assimilation of hexadecane-1-(14)C into cellular lipids is presented. Radioactivity in neutral lipid increased approximately sevenfold over the growth cycle, whereas radioactivity in phospholipid increased 50-fold during the same time period. The incorporation of radioactive fatty acids derived from the direct oxidation of hexadecane-1-(14)C demonstrated differential kinetics of assimilation into PE, PG, and CL. The results indicated a rapid turnover of phospholipid fatty acids in M. cerificans growing at the expense of hexadecane. 相似文献
11.
Ostrander DB Zhang M Mileykovskaya E Rho M Dowhan W 《The Journal of biological chemistry》2001,276(27):25262-25272
12.
A rapidly metabolizing pool of phosphatidylglycerol as a precursor of phosphatidylethanolamine and diglyceride in Bacillus megaterium. 总被引:5,自引:5,他引:0
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Pulse-chase experiments in Bacillus megaterium ATCC 14581 with [U-14C]palmitate, L-[U-14C]serine, and [U-14C]glycerol showed that a large pool of phosphatidylglycerol (PG) which exhibited rapid turnover in the phosphate moiety (PGt) underwent very rapid interconversion with the large diglyceride (DG) pool. Kinetics of DG labeling indicated that the fatty acyl and diacylated glycerol moieties of PGt were also utilized as precursors for net DG formation. The [U-14C]glycerol pulse-chase results also confirmed the presence of a second, metabolically stable pool of PG (PGs), which was deduced from [32P]phosphate studies. The other major phospholipid, phosphatidylethanolamine (PE), exhibited pronounced lags relative to PG and DG in 14C-fatty acid, [14C]glycerol, and [32P]phosphate incorporation, but not for incorporation of L-[U-14C]serine into the ethanolamine group of PE or into the serine moiety of the small phosphatidylserine (PS) pool. Furthermore, initial rates of L-[U-14C]serine incorporation into the serine and ethanolamine moieties of PS and PE were unaffected by cerulenin. The results provided compelling in vivo evidence that de novo PGt, PS, and PE synthesis in this organism proceed for the most part sequentially in the order PGt yields PS yields PE rather than via branching pathways from a common intermediate and that the phosphatidyl moiety in PS and PE is derived largely from the corresponding moiety in PGt, whereas the DG pool indirectly provides an additional source for this conversion by way of the facile PGt in equilibrium or formed from DG interconversion. 相似文献
13.
Absence of cardiolipin in the crd1 null mutant results in decreased mitochondrial membrane potential and reduced mitochondrial function 总被引:16,自引:0,他引:16
Jiang F Ryan MT Schlame M Zhao M Gu Z Klingenberg M Pfanner N Greenberg ML 《The Journal of biological chemistry》2000,275(29):22387-22394
Cardiolipin (CL) is a unique phospholipid which is present throughout the eukaryotic kingdom and is localized in mitochondrial membranes. Saccharomyces cerevisiae cells containing a disruption of CRD1, the structural gene encoding CL synthase, have no CL in mitochondrial membranes. To elucidate the physiological role of CL, we compared mitochondrial functions in the crd1Delta mutant and isogenic wild type. The crd1Delta mutant loses viability at elevated temperature, and prolonged culture at 37 degrees C leads to loss of the mitochondrial genome. Mutant membranes have increased phosphatidylglycerol (PG) when grown in a nonfermentable carbon source but have almost no detectable PG in medium containing glucose. In glucose-grown cells, maximum respiratory rate, ATPase and cytochrome oxidase activities, and protein import are deficient in the mutant. The ADP/ATP carrier is defective even during growth in a nonfermentable carbon source. The mitochondrial membrane potential is decreased in mutant cells. The decrease is more pronounced in glucose-grown cells, which lack PG, but is also apparent in membranes containing PG (i.e. in nonfermentable carbon sources). We propose that CL is required for maintaining the mitochondrial membrane potential and that reduced membrane potential in the absence of CL leads to defects in protein import and other mitochondrial functions. 相似文献
14.
Romantsov T Stalker L Culham DE Wood JM 《The Journal of biological chemistry》2008,283(18):12314-12323
The phospholipid composition of the membrane and transporter structure control the subcellular location and function of osmosensory transporter ProP in Escherichia coli. Growth in media of increasing osmolality increases, and entry to stationary phase decreases, the proportion of phosphatidate in anionic lipids (phosphatidylglycerol (PG) plus cardiolipin (CL)). Both treatments increase the CL:PG ratio. Transporters ProP and LacY are concentrated with CL (and not PG) near cell poles and septa. The polar concentration of ProP is CL-dependent. Here we show that the polar concentration of LacY is CL-independent. The osmotic activation threshold of ProP was directly proportional to the CL content of wild type bacteria, the PG content of CL-deficient bacteria, and the anionic lipid content of cells and proteoliposomes. CL was effective at a lower concentration in cells than in proteoliposomes, and at a much lower concentration than PG in either system. Thus, in wild type bacteria, osmotic induction of CL synthesis and concentration of ProP with CL at the cell poles adjust the osmotic activation threshold of ProP to match ambient conditions. ProP proteins linked by homodimeric, C-terminal coiled-coils are known to activate at lower osmolalities than those without such structures and coiled-coil disrupting mutations raise the osmotic activation threshold. Here we show that these mutations also prevent polar concentration of ProP. Stabilization of the C-terminal coiled-coil by covalent cross-linking of introduced Cys reverses the impact of increasing CL on the osmotic activation of ProP. Association of ProP C termini with the CL-rich membrane at cell poles may raise the osmotic activation threshold by blocking coiled-coil formation. Mutations that block coiled-coil formation may also block association of the C termini with the CL-rich membrane. 相似文献
15.
16.
Cardiolipin promotes polar localization of osmosensory transporter ProP in Escherichia coli 总被引:2,自引:1,他引:1
Romantsov T Helbig S Culham DE Gill C Stalker L Wood JM 《Molecular microbiology》2007,64(6):1455-1465
17.
Phospholipid metabolism during bacterial growth 总被引:27,自引:0,他引:27
Haemophilus parainfluenzae incorporates glycerol and phosphate into the membrane phospholipids without lag during logarithmic growth. In phosphatidyl glycerol (PG), the phosphate and unacylated glycerol moieties turn over and incorporate radioactivity much more rapidly than does the diacylated glycerol. At least half the radioactivity is lost from the phosphate and unacylated glycerol in about 1 doubling. The total fatty acids turn over slightly faster than the diacyl glycerol. In phosphatidyl ethanolamine (PE), which is the major lipid of the bacterium, ethanolamine and phosphate turn over and incorporate radioactivity at least half as fast as the phosphate in PG. The glycerol of PE did not turn over in 4 bacterial doublings. In phosphatidic acid the glycerol turns over at one-third the rate of phosphate turnover. By means of a modified method for the quantitative recovery of 1,3-glycerol diphosphate from cardiolipin, the phosphates and middle glycerol of cardiolipin were shown to turn over more rapidly than the acylated glycerols during bacterial growth. There is no randomization of the radioactivity in the 1- and 3-positions of the glycerol in the course of 1 doubling. The fatty acids of PG turn over faster than those in PE. In both lipids the 2-fatty acids turn over much faster than the 1-fatty acids. At both positions the individual fatty acids have their own rates of turnover. The distribution of fatty acids between the 1- and 2-positions is the same as in other organisms, with more monoenoic and long-chain fatty acids at the 2-position. The different rates of turnover and incorporation of radioactivity into different parts of the lipids suggest that exchange reactions may be important to phospholipid metabolism. 相似文献
18.
Production and Repair of Radiochemical Damage in Escherichia coli Deoxyribonucleic Acid; Its Modification by Culture Conditions and Relation to Survival 总被引:22,自引:15,他引:7
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Late log-phase Escherichia coli B/r cells are 1.6 times more sensitive to killing by X rays than are stationary-phase cells when grown in Brain Heart Infusion (BHI) + glucose. The number of single-chain breaks formed per krad is the same for log- and stationary-phase cells. Stationary-phase cells show a somewhat greater ability to repair single-chain breaks (especially after high doses of X rays) than do log-phase cells. The rapidity and extent of postirradiation deoxyribonucleic acid (DNA) degradation are greater in log-phase cells than in stationary-phase cells. The enhanced viability exhibited by stationary-phase cells thus appears to correlate both with enhanced single-chain break repair and the reduced degradation of DNA. Cells grown to stationary phase in peptone medium (PO cells) are 3.4 times more sensitive to killing by X rays than cells grown to stationary phase in peptone medium supplemented with glucose and phosphate buffer (PG cells). The yield of single-strand breaks is the same for both types of cells (but the absolute yield is about two times higher than in the cells grown in BHI + glucose). The kinetics for the repair of single-chain breaks are the same for both types of cells for about 30 min. After this time period, further repair ceases in the PO cells but continues in the PG cells, provided that glucose is present in the medium. Postirradiation DNA degradation is both more rapid and more extensive in PO cells than in PG cells whether or not glucose is present in the postirradiation incubation medium. The survival of stationary-phase E. coli B/r grown in PO or PG medium is likewise unaffected by the presence of glucose in the plating medium, and thus correlates better with the lower DNA degradation seen in the PG cells than with the increased strand rejoining, since this latter process requires the presence of glucose. 相似文献
19.
Metabolism of phosphatidylglycerol and bis(monoacylglycero)-phosphate in macrophage subcellular fractions 总被引:1,自引:0,他引:1
M Waite L King T Thornburg G Osthoff T Y Thuren 《The Journal of biological chemistry》1990,265(35):21720-21726
Bis(monoacylglycero)phosphate (BMP) is synthesized from exogenous phosphatidylglycerol (PG) by macrophages (Cochran, F. R., Roddick, V. L., Connor, J. R., Thornburg, J. T., and Waite, M. (1987) J. Immunol. 138, 1877-1883). Previous work from our laboratory showed that arachidonic acid in BMP was released by the macrophages upon challenge of the cells with PMA (Cochran, F. R., Connor, J. R., Roddick, V. L., and Waite, M. (1985) Biochem. Biophys. Res. Commun. 130, 800-806). Here we extend those studies using a model cultured cell line of macrophages, RAW 264.7. When PG labeled with 32P- and [3H]glycerol in both moieties was added to the culture medium, 32P/[3H]BMP was synthesized in a time-dependent manner. Fractionation of cell homogenates on a discontinuous sucrose gradient in which the light membranes were floated from dense sucrose showed an enrichment of [3H]BMP in light membrane fractions. The precursor [3H]PG was also found in the light fractions but, relative to the [3H]BMP, was more abundant in the denser membrane fractions. The appearance of [3H]PG and [3H]BMP in the light membrane fraction was time-dependent which suggested that the initial uptake and metabolism of [3H]PG was into the denser membranes. Incubation of the light membranes under conditions that are optimal for the lysosomal phospholipase A1 led to significant metabolism of [3H]PG. Both degradation of [3H]PG to water-soluble compounds and its conversion to acylphosphatidylglycerol occurred while no lyso-PG was detected. On the other hand, little BMP was found to be degraded. From these studies we postulate that in lysosomes acylphosphatidylglycerol is a precursor of BMP and that the previously reported turnover of arachidonic acid by BMP may occur via transacylation rather than hydrolysis. 相似文献
20.
Cardiolipin (CL) is a major mitochondrial membrane phospholipid in the mammalian heart and the remodeling of CL is essential to maintain its unique unsaturated fatty acyl composition. We examined CL de novo biosynthesis and remodeling in the surviving population of H9c2 cardiac myoblast cells exposed to 2-deoxyglucose (2-DG). H9c2 cells were incubated in the absence or presence of 2-DG for 16 h with [1,3-3H]glycerol or [1-14C]linoleic acid (bound to albumin in a 1:1 molar ratio). Dead cells were removed and radioactivity was incorporated into CL. Its pool size, fatty acid composition, and the activities of the CL biosynthesis and remodeling enzymes were determined. The CL pool size, its fatty acid composition, and [1,3-3H]glycerol or [1-14C]linoleic acid incorporated into CL were unaltered in the surviving population of 2-DG-treated cells compared with controls. In addition, the activities of the CL de novo biosynthetic enzymes were unaltered. Cleaved caspase-3 and poly(ADP-ribose) polymerase were slightly elevated in the surviving population of 2-DG-treated cells compared with controls, indicating that apoptosis induction was occurring in these cells. Mitochondrial phospholipase A2 and monolysocardiolipin acyltransferase (MLCL AT) activities increased 33% (p < 0.05) and 63% (p < 0.05), respectively, in 2-deoxyglucose-treated cells compared with controls. In contrast, the activity of ALCAT1, an endoplasmic reticulum MLCL AT, decreased 77% (p < 0.05), but this was not due to a reduction in ALCAT1 mRNA expression. The mRNA expression of the Barth syndrome gene TAZ, encoding a mitochondrial CL transacylase, was unaltered in 2-DG treated cells. The increase in mitochondrial MLCL AT activity was due to an elevated expression in MLCL AT protein. Thus, an increase in MLCL AT activity and expression occurs to maintain the CL pool in the surviving population of H9c2 cells as a compensatory mechanism for the elevated phospholipase A2 activity seen in 2-DG-induced apoptosis. We hypothesize that increased mitochondrial MLCL AT activity and its expression, and hence, elevated CL resynthesis, may be a protective mechanism against monolysocardiolipin-mediated apoptosis. 相似文献