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21.
Interaction of fatty acids with recombinant rat intestinal and liver fatty acid-binding proteins 总被引:9,自引:0,他引:9
G Nemecz T Hubbell J R Jefferson J B Lowe F Schroeder 《Archives of biochemistry and biophysics》1991,286(1):300-309
Intestinal enterocytes contain two homologous fatty acid-binding proteins, intestinal fatty acid-binding protein (I-FABP)2 and liver fatty acid-binding protein (L-FABP). Since the functional basis for this multiplicity is not known, the fatty acid-binding specificity of recombinant forms of both rat I-FABP and rat L-FABP was examined. A systematic comparative analysis of the 18 carbon chain length fatty acid binding parameters, using both radiolabeled (stearic, oleic, and linoleic) and fluorescent (trans-parinaric and cis-parinaric) fatty acids, was undertaken. Results obtained with a classical Lipidex-1000 binding assay, which requires separation of bound from free fatty acid, were confirmed with a fluorescent fatty acid-binding assay not requiring separation of bound and unbound ligand. Depending on the nature of the fatty acid ligand, I-FABP bound fatty acid had dissociation constants between 0.2 and 3.1 microM and a consistent 1:1 molar ratio. The dissociation constants for L-FABP bound fatty acids ranged between 0.9 and 2.6 microM and the protein bound up to 2 mol fatty acid per mole of protein. Both fatty acid-binding proteins exhibited relatively higher affinity for unsaturated fatty acids as compared to saturated fatty acids of the same chain length. cis-Parinaric acid or trans-parinaric acid (each containing four double bonds) bound to L-FABP and I-FABP were displaced in a competitive manner by non-fluorescent fatty acid. Hill plots of the binding of cis- and trans- parinaric acid to L-FABP showed that the binding affinities of the two sites were very similar and did not exhibit cooperativity. The lack of fluorescence self-quenching upon binding 2 mol of either trans- or cis-parinaric acid/mol L-FABP is consistent with the presence of two binding sites with dissimilar orientation in the L-FABP. Thus, the difference in binding capacity between I-FABP and L-FABP predicts a structurally different binding site or sites. 相似文献
22.
Altered membrane structure in transfected mouse L-cell fibroblasts expressing rat liver fatty acid-binding protein 总被引:7,自引:0,他引:7
J R Jefferson D M Powell Z Rymaszewski J Kukowska-Latallo J B Lowe F Schroeder 《The Journal of biological chemistry》1990,265(19):11062-11068
Mouse L cell fibroblasts were transfected with cloned cDNA encoding rat liver fatty acid binding protein (L-FABP) also known as sterol carrier protein. Stable transfectant cell lines were selected and expression of L-FABP determined using Western blot analysis. The nontransfected controls and low expression cells did not differ significantly in any of the properties examined. All cell lines showed similar doubling times but cells expressing high levels of L-FABP attained 2-fold higher cell saturation density and differed significantly in their lipid metabolism as indicated by 1) higher cholesterol ester and phospholipid content, and 2) decreased sterol/phospholipid ratio. The observed changes in the lipid composition predicted a lower degree of membrane-lipid order (higher fluidity) in the plasma membranes of cells expressing high levels of L-FABP. Therefore, fluorescent molecule, 1,6-diphenyl-1,3,5-hexatriene, and multifrequency (1-250 MHz) phase and modulation fluorometry were used to probe the effect of L-FABP expression on membrane structure. Steady-state polarization and limiting anisotropy of diphenylhexatriene were significantly lower in the isolated plasma membrane vesicles from the high expression clones. The observed changes in L-cells as a result of de novo expression of L-FABP are consistent with the ability of this protein to bind sterols and fatty acids, stimulate sterol esterification, and stimulate phospholipid biosynthesis. This evidence is supportive of a physiologic role for L-FABP in modulating cellular lipid metabolism and membrane structure. 相似文献
23.
Robert L. Wykle Craig H. Miller Jon C. Lewis Jeffrey D. Schmitt Jennie A. Smith Jefferson R. Surles Claude Piantadosi Joseph T. OFlaherty 《Biochemical and biophysical research communications》1981,100(4):1651-1658
1-O-Hexadecyl-2-O-acetyl--glycero-3-phosphocholine (platelet activating factor) stimulated the degranulation of rabbit platelets and human neutrophils, whereas the enantiomer, 3-O-hexadecyl-2-O-acetyl--glycero-1-phosphocholine, was inactive. The analogs compared had the following relative potencies in degranulating platelets and neutrophils: 1-O-hexadecyl-2-O-acetyl--glycero-3-phosphocholine > 1-O-hexadecyl-2-O-ethyl--glycero-3-phosphocholine >-1-O-octadecyl-2-O-ethylglycero-3-phosphocholine = 1-O-hexadecyl-2-O-methyl--glycero-3-phosphocholine >-1-O-dodecyl-2-O-ethyl-glycero-3-phosphocholine. The deacetylated compound, 1-O-hexadecyl-2-lyso--glycero-3-phosphocholine, and 1-O-hexadecyl-2,2-dimethylpropanediol-3-phosphocholine were inactive. The active analogs selectively desensitized the response to each other in the neutrophils. It is suggested that these compounds may activate cells through interaction with a stereospecific receptor. 相似文献
24.
Reproductive ecology of Naididae (Oligochaeta) 总被引:5,自引:5,他引:0
Jefferson Parish 《Hydrobiologia》1981,83(1):115-123
Asexual reproduction is employed by species of Naididae during favorable environmental conditions. In species characteristic of aquatic habitats subject to rapid fluctuations in water levels and temperatures most individuals in a population become sexually mature, and there is degeneration of the alimentary tract, a shortened breeding season, and aclitellar formation of cocoons. Cocoon deposition allows a population to survive periods of environmental stress. Sexual reproduction predominates near the edges of ranges where stress conditions are more prevalent. Species that reproduce sexually in spring are intolerant of summer conditions, while those that breed in autumn are intolerant of winter. Species typical of environmentally stable bodies of water usually have a minority of the population mature at any time, an extended breeding season, and the ability to continue feeding while mature; thus, adaptations for survival in unstable habitats are lacking. 相似文献
25.
Elizabeth Witkoski Mary F. Schuler Richard C. Feldhoff Samson I. Jacob Leonard S. Jefferson Allan Lipton 《Experimental cell research》1979,124(2)
Isolated rat hepatocytes release an acidic glycoprotein(s) that can selectively promote the growth of transformed cells. This factor has a molecular weight of 60 000–70 000 D. Liver microsomal and cytosol fractions contain two species of stimulatory activity—44 000 and 3 500 D. Mitochondrial and nuclear fractions contain only the lower molecular weight factor. 相似文献
26.
The effects of growth hormone, administered in vivo or added in vitro, on amino acid transport and utilization have been studied in perfused livers of normal and hypophysectomized rats. A perfusion system employing a nonrecirculating medium was used in all of the studies. Two nonmetalbolizable amino acid analogues, alpha-aminoisobutyric acid (AIB) and 1-aminocyclopentane carboxylic acid (cycloleucine) were used to study transport. Accumulation of AIB increased linearly over a 60-min perfusion period, reaching distribution ratios of between 1 and 2 for both groups of animals. Treatment of both normal and hypophysectomized rats with growth hormone 60 min prior to the start of perfusion increased AIB distribution ratios by up to 84 and 108%, respectively. Accumulation of cycloleucine was linear for only about 20 min of perfusion and then plateaued. Steady state distribution ratios of this analogue ranged between 1 and 2 for both groups of animals. Growth hormone treatment had no apparent effect on the time necessary to reach these steady state levels, but significantly increased them in livers of both normal and hypophysectomized rats by 16 and 42%, respectively. Studies designed to analyze the kinetic properties of these hormone effects revealed that growth hormone treatment caused 2-fold i-crease in the maximum velocities of both the AIB and cycloleucine transport systems. The substrate concentration for half-maximal transport velocity was increased slightly for both systems by growth hormone. Direct effects of growth hormone were demonstrated in studies where livers of hypophysectomized rats were perfused under conditions simulationg those of experiments in which the hormone was administered in vivo. Following an initial 45-min period of perfusion the medium during the 20 min. Growth hormone added to the medium during the entire 65-min perfusion at a concentration of 1 mug per ml caused a 30% increase in the cycloleucine distribution ratio. Under similar experimental conditions growth hormone directly stimulated three hepatic pathways of amino acid utilization: (a) incorporation of [14C]valine into protein, (b) urea formation and (c) conversion of 14-C-amino-acids to labeled glucose. Intracellular concentrations of seven amino acids, including threonine, serine, proline, glycine, alanine, lysine, and arginine, were increased significantly in livers perfused with medium containing growth hormone... 相似文献
27.
Adiantum diphyllum, a species endemic to the coastal forests of Bahia State, Brazil, is characterized and distinguished from other species ofAdiantum with anastomosing veins, elongate sori, and venuloid idioblasts.
Adiantum diphyllum, uma espécie endêmica das florestas costeiras do leste do Brasil. é caracterizada e distinta das outras espécies deAdiantum com nervuras anastomosadas, soros alongados e idioblastos entre as nervuras.相似文献
28.
S R Kimball D A Antonetti R M Brawley L S Jefferson 《The Journal of biological chemistry》1991,266(3):1969-1976
In previous studies, initiation of protein synthesis was shown to be inhibited in perfused rat livers deprived of single essential amino acids. In the present study, histidinol, a competitive inhibitor of histidinyl-tRNA synthetase, was used to amplify the effects of histidine deprivation on protein synthesis in perfused liver to facilitate investigation of mechanisms involved in the inhibition of peptide chain initiation. Protein synthesis was reduced to 77% of the control rate in livers deprived of histidine and to 13% of the control rate in livers deprived of histidine and exposed to 2.0 mM histidinol. The inhibition of protein synthesis caused by histidine deprivation alone was accompanied by a 2-fold increase in the number of free ribosomal particles, a 29% decrease in Met-tRNA(i) binding to 43 S preinitiation complexes, and a 31% reduction in activity of eukaryotic initiation factor 2B (eIF-2B). By comparison, histidine deprivation combined with histidinol addition resulted in a 3-fold increase in free ribosomal particles, a 66% decrease in Met-tRNAi binding, and a 78% reduction in eIF-2B activity. The proportion of the alpha-subunit of eukaryotic initiation factor two (eIF-2) in the phosphorylated form increased from 8.9 +/- 0.8% in control livers to 52.4 +/- 5.5% in response to histidinol. The increase in the amount of eIF-2 alpha in the phosphorylated form apparently was not due to an increase in kinase activity, because there was no change in eIF-2 alpha kinase activity in extracts of liver perfused with medium containing histidinol compared to controls. Instead, the increased phosphorylation of eIF-2 alpha was associated with an inhibition of eIF-2 alpha phosphatase activity. Thus, in contrast to other systems that have been examined, the mechanism involved in the increase in the phosphorylation state of eIF-2 alpha appears to involve an inhibition of eIF-2 alpha phosphatase activity rather than activation of an eIF-2 alpha kinase. 相似文献
29.
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