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M K Wassef

Publications and source records attributed to M K Wassef.

14 recordsLinked to original sources

Lipid analyses of isolated surface membranes of Leishmania donovani promastigotes.

Constituent lipids of surface membranes (SM) isolated from Leishmania donovani promastigotes were analyzed and compared with those obtained from whole cells and an isolated kinetoplast-mitochondrion fraction (KM). On a dry weight basis, the total extractable lipids constituted approximately 47%, 12% and 24% of the SM, cells and KM, respectively. The total lipids of SM, cells and KM all were composed of approximately 70% phospholipids (PL), 20-25% neutral lipids and 5-10% glycolipids. Sterols and diglycerides composed 60% and 30%, respectively, of the various neutral lipid fractions. Several mannose- and galactose-containing glycolipids were fractionated but not identified. The glycolipid fractions from cells and SM had demonstrable antigenic activities with rabbit anti-SM sera. Striking quantitative differences were apparent between the PL profiles of the 3 cellular components examined. The PL of SM, whole cells and KM, respectively, were composed of: 15%, 51% and 24% phosphatidylcholine; 37%, 13% and 11% phosphatidylethanolamine (PE); 18%, 10% and 14% phosphatidylinositol; 10%, 1% and 4% phosphatidylserine and traces of cardiolipin, phosphatidylglycerol and phosphatidic acid. An unknown PL containing sphingosine, choline and vicinal hydroxyl groups but no free amino moieties made up approximately 19% of the PL of SM and whole cells, but it constituted approximately 27% of the PL of KM. The PL side chain constituents of whole cells and SM were composed mainly of longchain fatty acids (C18-20). Further, over 50% of the PE of SM was in the alkyl and alK-1-enyl ether forms. These SM properties might contribute to the organism's resistance to digestion in the hydrolytic environs of both its insect vector and mammalian hosts.

Animals↗

Degradation of phosphatidylinositol by soluble enzymes of rat gastric mucosa.

Rat gastric mucosa homogenates contain two enzymatic systems for hydrolyzing phosphatidylinositol: a deacylation activity yielding lysophosphatidylinositol and free fatty acid, and a phospholipase C-like activity producing 1,2-diacylglycerol and inositol phosphates. These activities were found mainly in the 105 000 x g supernatant and could be distinguished by differential stabilities, metal requirements and the action of deoxycholate and mepacrine. Each lipolytic reaction displayed a major pH optimum at 7.5 and a minor pH optimum at 5.5. The deacylation system was 8-10 times as active as the phospholipase C, with an apparent Km of 0.63 mM towards 1-acyl-2-arachidonylphosphatidylinositol at pH 7.5. The phospholipase C activity, on the other hand, hydrolyzed 1-acyl-2-arachidonylphosphatidylinositol or 1-acyl-2-arachidonylphosphatidylethanolamine and yielded 1-acyl-2-arachidonyl-sn-glycerol. This 1,2-diacylglycerol could be phosphorylated to form 1-acyl-2-[14C]arachidonyl-sn-phosphoglycerol (phosphatidic acid), but could not be hydrolyzed to produce free [14C]arachidonic acid using stomach mucosal microsomes. Phospholipase A2 and phospholipase C attack 1-acyl-2-arachidonylphosphatidylethanolamine and phosphatidylinositol equally well, but hydrolyze 1-acyl-2-arachidonylphosphatidylcholine poorly.

Animals↗

Molecular species of phosphatidylcholine from rat gastric mucosa.

Phosphatidylcholine of rat gastric mucosa were found to constitute about half of the total phospholipids. The composition of 20 molecular species accounting for approx. 90% of the total phosphatidylcholine was determined by specific enzymic hydrolyses and AgNO3 thin-layer and gas-liquid chromatography. Disaturated (dipalmitoyl) phosphatidylcholine made up about 31% of the total phosphatidylcholines. Other species which occurred in significant concentrations included 16:0/18:1, 18:0/18:1, 16:0/18:2, 18:0/18:2, 16:0/18:3, 18:0/18:3, 16:0/20:4, and 18:0/20:4. These results indicate that rat gastric mucosa is similar to lung in that both contain elevated amounts of dipalmitoyl phosphatidylcholine. Other similarities between these two tissues are discussed.

Animals↗

Phospholipid-deacylating enzymes of rat stomach mucosa.

1. Rat stomach mucosa exhibited three distinguishable phospholipid-deacylating enzyme activities: lysophospholipase, phospholipase A1 and phospholipase A2. 2. The lysophospholipase hydrolyzed 1-palmitoyl lysophosphatidylcholine to free fatty acid and glycerophosphorylcholine. This enzyme had an optimum pH of 8.0, was heat labile, did not require Ca2+ for maximum activity and was not inhibited by bile salts or buffers of high ionic strength. 3. Phospholipase A2 and phospholipase A1 deacylated dipalmitoyl phophatidylcholine to the corresponding lyso compound and free fatty acid. The specific activity of phospholipase A2 was 2--4-fold higher than that of phospholipase A1 under all the conditions tested. Both activities were enhanced 4--7.5-fold in the presence of bile salts at alkaline pH and 11-18-fold at acidic pH. 4. In the absence of bile salts, phospholipase A1 exhibited pH optima at 6.5 and 9.5 and phospholipase A2 at pH 6.5, 8.0 and 9.5. The pH optima for phospholipase A1 were shifted to pH 3.0, 6.0 and 9.0 in presence of sodium taurocholate; the activity was detected only at a single pH of 9.5 in the presence of sodium deoxycholate and at pH 10.0 in the presence of sodium glycocholate. Phospholipase A2 optimum activity was displayed at pH 3.0, 6.0 and 8.0 in presence of taurocholage, pH 7.5 and 9.0, in presence of glycocholate and only at pH 9.0 in presence of deoxycholate. 5. Ca2+ was essential for optimum activity of phospholipases A1 and A2. But phospholipase A1 lost complete activity in presence of 0.5 mM ethyleneglycolbis-(beta-aminoethylether)-N,N'-tetraacetic acid (EGTA) at pH 6.0, whereas phospholipase A2 lost only 50%. 6. Phospholipases A1 and A2 retained about 50% of their activities by heating at 75 degrees for 10 min. At 100 degrees, phospholipase A1 retained 22% of its activity, whereas phospholipase A2 retained only 7%.

Animals↗

Ceramide aminoethylphosphonate in the fungus Pythium prolatum.

Ceramide aminoethylphosphonate was characterized from the lower fungus Pythium prolatum. The compound was purified by silicic acid column chromatography, DEAE-cellulose column chromatography, the action of phospholipase D, and two-dimensional thin-layer chromatography. Infrared spectra lacked ester bands and suggested the presence of a bonded NH group. The compound was hydrolyzed by strong acid. Sphingosine comprised 98% of the long chain bases. The predominant fatty acids were palmitate, oleate, linoleate, and an unidentified long chain acid. The aqueous portion of the hydrolysis products gave an elemental analysis consistent with 2-aminoethylphosphonate. On paper and thin layer chromatograms, the ammonium salt of the aqueous hydrolysis product chromatographed with 2-aminoethylphosphoate but not 1-aminoethylphosphonate, 2-aminoethanol, serine or alanine. This appears to be the first report of a phosphonolipid from fungi.

Aminoethylphosphonic Acid↗

Lipids of Klebsiella pneumoniae: the presence of phosphatidyl choline in succinate-grown cells.

The carbon and energy source for aerobically grown cultures of Klebsiella pneumoniae profoundly influenced the total lipid content and phosphatide composition. Glucose-grown cells contained 13% lipid, 56% of which was phospholipids. Succinate-grown cells contained 8% lipid, 66% of which was phospholipids. The predominant phosphatides of glucose-grown cells were phosphatidyl ethanolamine, 82%; phosphatidyl glycerol, 4.5%; phosphatidic acid, 5%; cardiolipin, 6.5%; phosphatidyl serine; and trace amounts of unidentified phosphatides. Phosphatides of succinate-grown cells were phosphatidyl ethanolamine, 38%; diphosphatidyl glycerol, 14%; phosphatidyl glycerol, 13%; phosphatidyl choline, 14.5%; phosphatidyl serine, 6%; phosphatidic acid, 4%; and 10% unknown lipids. No trace of phosphatidyl choline was found in glucose-grown cells.

Chromatography↗

Lipid chemistry.

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Alcohols↗