Utilization of fatty acid supplements by cultured animal cells

Williams, RE; Bj, Wisnieski; Hg, Rittenhouse; Cf, Fox

doi:10.1021/bi00706a029

Cited by 82 publications

(22 citation statements)

References 18 publications

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“…The phospholipids of other cultured mammalian cell types have been enriched for saturated or unsaturated fatty acids by various techniques (15)(16)(17), with effects on such membrane properties as concanavalin A binding (18) and lectin-induced cell agglutination (16) which correlated with differences induced in saturated fatty acid content. Similarly, a report concerning nonreceptor-mediated phagocytosis in thioglycollate-stimulated peritoneal macrophages incubated for 4 days in medium containing individual fatty acids appeared during the course of our work and showed reduced particle uptake at 370 (19).…”

Section: Methodsmentioning

confidence: 99%

Response of endocytosis to altered fatty acyl composition of macrophage phospholipids.

Mahoney¹,

Hamill²,

Scott³

et al. 1977

Proc. Natl. Acad. Sci. U.S.A.

140

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Mouse peritoneal macrophages incubated in serumless medium containing a 19:0 or trans-18:1 fatty acid complexed to bovine serum albu'min incorporate the exogenous fatty acid supplement into cellular phospholipids. Within 8 hr, 25% of the total phospholipid fatty acids are derived from the supplement, with cell viability remaining >95%. The incorporation of either of these supplements increases the saturated/ unsaturated fatty acid ratio in the phospholipids 2-fold over that of cells cultured in serum and effects striking changes in endocytic activities. The levels of both fluid-phase pinocytosis and receptor-mediated phagocytosis are decreased at all temperatures examined between 150 and 37°. The increased degree of saturation of cell phospholipids correlates with decreased endocytic rates for both processes and with increased activation energies (Eact) for phagocytosis. The Eact values for phagocytosis, which range from 54 to 90 kcal/mol, depend on the supplementation conditions used. Although the levels of pinocytosis are depressed, the Ea. values for pinocytosis (17-25 kcal/mol)

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Section: Methodsmentioning

confidence: 99%

Response of endocytosis to altered fatty acyl composition of macrophage phospholipids.

Mahoney¹,

Hamill²,

Scott³

et al. 1977

Proc. Natl. Acad. Sci. U.S.A.

140

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“…reconstitution studies is the modification of the membrane lipid composition in vivo. Brivio-Haugland et al showed significant changes in basal and hormone-stimulated adenylate cyclase activity when rats were fed an essential fatty acid deficient diet (18 (20)(21)(22)(23)(24) and polar head group composition (24,25). These developments now make it possible to carry out enzymatic and physical studies in the membranes of animal cells with biosynthetically modified lipid compositions (22,23,26).…”

mentioning

confidence: 99%

Modification of adenylate cyclase activity in LM cells by manipulation of the membrane phospholipid composition in vivo.

Engelhard¹,

Storm²,

Gläser³

1976

Proc. Natl. Acad. Sci. U.S.A.

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Adenylate cyclase [ATP pyrophosphate-lyase (cyclizing); EC 4.6.1.1] activities were examined in mouse LM cell (fibroblast) membranes that were supplemented with ethanolamine and/or fatty acids. The supplements were incorporated into the plasma membrane phospholipids in significant amounts. Fatty acid supplementations had distinct effects as compared to polar head group supplementations. All lipid supplementations increased basal adenylate cyclase activity re ative to control cells grown in choline-containing medium. Double supplementation with ethanolamine and linoleate increased the specific activity of adenylate cyclase up to 4-fold. Activity in the presence of fluoride was unaffected by ethanolamine supplementation, but was increased by fatty acid supplementation. In contrast, prostaglandin E1 stimulation was 4.2-fold in controls and ethanolamine and/or elaidate supplements, 6-fold in choline plus linoleate supplements, and 3.1-told in ethanolamine plus linoleate supplements. Differences in activity could not be ascribed to changes in membrane protein composition in supplemented cells, and could be abolished by detergent solubilization. The fluidity of the supplemented membranes was monitored by fluorescence polarization, and no correlation was observed between membrane viscosity and adenylate cyclase activity or hormone stimulation. These results emphasize the importance of the membrane lipid phase for this enzyme. Adenylate cyclase [ATP pyrophosphate-lyase (cyclizing); EC 4.6.1.1] is associated with the plasma membranes of animal cells (1-7). The enzyme is tightly bound to the membrane, and has been solubilized only by the use of membrane-disruptive agents such as detergents. Several lines of evidence indicate that adenylate cyclase is dependent upon membrane lipids for basal and hormone-stimulated activities. The membrane-bound enzyme has been treated with filipin (3), nonionic detergents (8-10), digitonin (11), phospholipases (11)(12)(13), and organic solvents (13). These treatments resulted in changes in either basal activity or hormone stimulation or both. When phospholipids were added back (11,(13)(14)(15)(16) Growth and Supplementation of Cells. Mouse LM cells were grown in suspension culture in Higuchi's medium (24) containing 20 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (Hepes), pH 7.4, 1 g/liter of methylcellulose, and 0.02 g/liter of sodium dextran sulfate. Growth and supplementation were carried out as described (20,24).Lipid Determinations. Lipids were extracted by the method of Bligh and Dyer (32) as described by Ames (33). Phospholipids were separated by two-dimensional thin-layer chromatography (20,24). Spots were visualized with I2 vapor, scraped, and eluted with 5 ml of CHCl3:CH30H:acetic acid:H20 (5:5:1:1, vol/vol) followed by 2 ml of CH30H. The extracts were combined, evaporated to dryness, and redissolved in CHCl3:CH30H:H20 (2:2:1.8, vol/vol

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“…The procedure used to modify the fatty acid composition of LM cells has been described (20). Confluent cells were detached from Falcon tissue flasks by treatment with 0.005% trypsin (Sigma: type 2, pancreatin; 1,500 BAEE units per mg) and seeded at an approximate 20-fold lower cell density on Linbro multidish trays (35 mm diameter).…”

Section: Methodsmentioning

confidence: 99%

“…Fatty acid analysis of the combined phosphatidylcholine (PC) and phosphatidylethanolamine (PE) fraction derived from cells grown in medium containing Tween-19:O at 8 pg/ml (fae) revealed a saturated fatty acid content of approximately 45% (20). In contrast, cells grown in MEM + P (control cells) contained about 30% saturated fatty acid in PC and PE.…”

Section: Effects Of Lipid Composition On Cell-lectin Interactionsmentioning

confidence: 99%

Effect of membrane lipid composition and microtubule structure on lectin interactions of mouse lm cells

Rittenhouse¹,

Williams²,

Fox³

1974

J Supramol Struct

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Concanavalin A (Con A) binding and Con A-mediated hemadsorption t o LM cells were found t o decrease significantly at both 5-7'C and 15-19'C. The higher of these critical temperatures responds t o a change in state of the membrane lipids and can be increased or decreased in cells where the membrane phospholipids contain less or more double bonds, respectively. The lower critical temperature for Con A binding or Con A-mediated hemadsorption does not respond t o these changes in membrane lipid composition. Though the amount of Con A bound t o the cell surface is a determinant of Con A-mediated agglutinability, the major components of the decreases in Con A-mediated hemadsorption which occur at both these critical temperatures do not have their origin in the decreases ip Con A bindi!g which occur over these same temperature ranges -that is 5-7 C and 15-19 C. , Con A-mediated hemadsorption measured at 22OC was dramatically inhibited when LM cells were first incubated at 7'C or less. Reversal of this inhibition required 20-30 min of subsequent incubation at 22"C, indicating that factors other than membrane lipid "fluidity" are determinants of agglutinability. LM cells treated with the microtubule-disrupting alkaloids colchicine, colcemid, or vinblastine at concentrations as low as agglutinable with Con A. By contrast, lumicolchicine, an inactive derivative of colchicine, had a slight inhibitory effect on Con A-mediated hemadsorption. Colchicine, vinblastine, or lumicolchicine treatment of LM cells did not alter the quantitative binding of labeled lectin. The results suggest that membrane lipid "fluidity" and the cell cytoskeleton (microtubule/microfilament system) are important determinants of lectin interactions with cell surfaces. The results are interpreted in terms of a model of cell-cell and cell-lectin interactions which assigns a central role t o the Con A receptor.

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Utilization of fatty acid supplements by cultured animal cells

Cited by 82 publications

References 18 publications

Response of endocytosis to altered fatty acyl composition of macrophage phospholipids.

Response of endocytosis to altered fatty acyl composition of macrophage phospholipids.

Modification of adenylate cyclase activity in LM cells by manipulation of the membrane phospholipid composition in vivo.

Effect of membrane lipid composition and microtubule structure on lectin interactions of mouse lm cells

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