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71.
The Lipid Peroxidation Product, 4-Hydroxy-2-trans-Nonenal, Alters the Conformation of Cortical Synaptosomal Membrane Proteins 总被引:6,自引:1,他引:5
‡Ram Subramaniam ‡Fred Roediger Brad Jordan †‡§Mark P. Mattson §Jeffrey N. Keller Georg Waeg ‡§ D. Allan Butterfield 《Journal of neurochemistry》1997,69(3):1161-1169
Abstract: Alzheimer's disease (AD) is widely held to be a disorder associated with oxidative stress due, in part, to the membrane action of amyloid β-peptide (Aβ). Aβ-associated free radicals cause lipid peroxidation, a major product of which is 4-hydroxy-2- trans -nonenal (HNE). We determined whether HNE would alter the conformation of synaptosomal membrane proteins, which might be related to the known neurotoxicity of Aβ and HNE. Electron paramagnetic resonance spectroscopy, using a protein-specific spin label, MAL-6(2,2,6,6-tetramethyl-4-maleimidopiperidin-1-oxyl), was used to probe conformational changes in gerbil cortical synaptosomal membrane proteins, and a lipid-specific stearic acid label, 5-nitroxide stearate, was used to probe for HNE-induced alterations in the fluidity of the bilayer domain of these membranes. Synaptosomal membranes, incubated with low concentrations of HNE, exhibited changes in protein conformation and bilayer order and motion (fluidity). The changes in protein conformation were found to be concentration- and time-dependent. Significant protein conformational changes were observed at physiologically relevant concentrations of 1–10 µ M HNE, reminiscent of similar changes in synaptosomal membrane proteins from senile plaque- and Aβ-rich AD hippocampal and inferior parietal brain regions. HNE-induced modifications in the physical state of gerbil synaptosomal membrane proteins were prevented completely by using excess glutathione ethyl ester, known to protect neurons from HNE-caused neurotoxicity. Membrane fluidity was found to increase at higher concentrations of HNE (50 µ M ). The results obtained are discussed with relevance to the hypothesis of Aβ-induced free radical-mediated lipid peroxidation, leading to subsequent HNE-induced alterations in the structure and function of key membrane proteins with consequent neurotoxicity in AD brain. 相似文献
72.
Complete nucleotide sequence of a gene encoding a functional human class I histocompatibility antigen (HLA-CW3) 总被引:21,自引:1,他引:20
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The HLA-CW3 gene contained in a cosmid clone identified by transfection expression experiments has been completely sequenced. This provides, for the first time, data on the structure of HLA-C locus products and constitutes, together with that of the gene coding for HLA-A3, the first complete nucleotide sequences of genes coding for serologically defined class I HLA molecules. In contrast to the organisation of the two class I HLA pseudogenes whose sequences have previously been determined, the sequence of the HLA-CW3 gene reveals an additional cytoplasmic encoding domain, making the organisation of this gene very similar to that of known H-2 class I genes and also the HLA-A3 gene. The deduced amino acid sequences of HLA-CW3 and HLA-A3 now allow a systematic comparison of such sequences of HLA class I molecules from the three classical transplantation antigen loci A, B, C. The compared sequences include the previously determined partial amino acid sequences of HLA-B7, HLA-B40, HLA-A2 and HLA-A28. The comparisons confirm the extreme polymorphism of HLA classical class I molecules, and permit a study of the level of diversity and the location of sequence differences. The distribution of differences is not uniform, most of them being located in the first and second extracellular domains, the third extracellular domain is extremely conserved, and the cytoplasmic domain is also a variable region. Although it is difficult to determine locus-specific regions, we have identified several candidate positions which may be C locus-specific. 相似文献
73.
The 135-nucleotide-long “5 + S” RNA molecule found in Drosophila tissue culture cells after labelling at 37 °C has been identified as a precursor to 5 S RNA by pulse-chase experiments. The structure of the 15-nucleotide-long 3′-terminal sequence which differentiates this molecule from mature 5 S RNA has been determined. This ends in a stretch of U residues, suggestive of a polymerase termination signal. 相似文献
74.
Summary Fox and Woese (1975a) have shown that a model of 5S RNA secondary structure similar to the one originally derived forChlorella 5S RNA can be generalized with relatively minor variations to all sequenced 5S RNA molecules, i.e. that corresponding base paired regions can be formed at approximately the same positions. We present experimental data in favour of this hypothesis and show that the points at which ribonucleases T1, T2 and pancreatic ribonuclease cleave six different 5S RNA molecules under mild conditions (high ionic strength, low temperature, low RNAase concentration) nearly always fall in the proposed single-stranded regions. We conclude that this model is a good approximation to the conformation of 5S RNA in solution. 相似文献
75.
Summary Intermolecular transposition of Tn2660 into pCR1 was measured at 30°C in recA
– and recA
+ hosts as between 2.6 and 5.5x10–3, a similar value to that previously found for Tn3. No cointegrate structures were found under conditions where 104 transposition events occurred. Immunity to intermolecular transposition of Tn2660, similar to that found for Tn3 was demonstrated by showing that the above transposition frequency was reduced by a factor of between 10–3 and 10–4 when a mutant Tn2660 (resulting in the synthesis of a temperaturesensitive -lactamase) was present in the recipient plasmid. Intramolecular transposition of Tn3 was found to occur under the same conditions as previously demonstrated for Tn2660 giving rise to similar end products, in which the newly introduced Tn3 is oriented inversely to the resident Tn3 and the DNA sequence between the two transposons has been inverted. Thus, in all respects functional identity of the transposition activities of Tn3 and Tn2660 is shown, thereby identifying characteristics of intramolecular transposition that are not readily accommodated by current models of transposition. 相似文献
76.
Summary A simple container is described whereby small seedlings may be grown at controlled levels of water stress. The water stress was induced in the soil by an osmoticum which is separated from the soil by a semi-permeable membrane. The mortality ofBetula pendula seedlings was markedly increased at a matric potential of –1.6 bars whereas the mortality ofBuddleia davidii was only affected below –2.8 bars. This difference in tolerance to water stress at the seedling stage might not be reflected in the distribution of the species in the colonisation of chalk and sand pits in England unless there is a dry spring. 相似文献
77.
C. Jordan W. Caskey G. Escalante R. Herrera F. Montagnini R. Todd C. Uhl 《Plant and Soil》1982,67(1-3):325-332
Standing stocks and fluxes of nitrogen, including nitrogen fixation and denitrification, were measured in a tropical rainforest on Oxisol in the Amazon Territory of Venezuela. The standing stock of nitrogen was comparable to that of temperate forests, but was higher than that in an adjacent forest on Spodosol. Fluxes were higher than in forests in the temperate zone, but lower than in another tropical forest on more fertile soil. Even though nitrogen was abundant, this does not mean that nitrogen could not be limiting to agriculture if the forest is cleared and the land cultivated. The nitrogen fixing and nitrogen conserving mechanisms are dependent upon the structure of the undisturbed forest, and destruction of the forest would eventually decrease the input of nitrogen to the soil. 相似文献
78.
Movement of Kinetin and Gibberellic Acid in Leaf Petioles during Water Stress-induced Abscission in Cotton
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Movement of [14C]kinetin and [14C]gibberellic acid was examined in cotton (Gossypium hirsutum L.) cotyledonary petiole sections independent of label uptake or exit from the tissue. Sections 20 millimeters in length were taken from well watered, stressed, and poststressed plants. Transport capacity was determined using a pulse-chase technique. Movement of both kinetin and gibberellic acid was found to be nonpolar with a velocity of 1 millimeter per hour or less, suggesting passive diffusion. Neither water stress nor anaerobic conditions during transport of labeled material affected the transport capacity of the petioles. 相似文献
79.
Control of porphyrin biosynthesis in Rhodopseudomonas spheroides and Propionibacterium shermanii. A direct 13C nuclear-magnetic-resonance spectroscopy study.
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G Burton P M Jordan N E MacKenzie P E Fagerness A I Scott 《The Biochemical journal》1981,194(2):627-631
The facultative anaerobes Rhodopseudomonas spheroides and Propionibacterium shermanii were grown under anaerobic and aerobic conditions. The effect of light was studied with the photosynthetic R. spheroides, and the adaptation of both species to dark anaerobic life was monitored by direct observation of 5-amino[5-13C]laevulinic acid metabolism by using 13C nuclear-magnetic-resonance spectroscopy. 相似文献
80.
A comparative study was carried out in order to determine which of the most commonly used alkalies for protein hydrolysis in tryptophan determination gave the best results. Hydrolyses were performed with 2.5 and 4 n Ba (OH)2, 4 and 10 n NaOH, 5 n NaOH containing 5% SnCl2, and with 4 n LiOH, not previously reported for use. The effect of temperature and hydrolysis time on the measured tryptophan content was also determined. Based on results obtained with lysozyme and with seven high protein preparations 4 n LiOH gave the best results. A temperature of 145°C was selected as the most convenient temperature since maximum tryptophan values were obtained with 4–8 h. The hydrolysis time required was inversely related to the protein content of the preparation. Lysozyme, casein, bovine plasma protein, and dehydrated whole egg gave maximum tryptophan content after 4 h hydrolysis while skimmed milk powder, rice flour, wheat flour, and wild legume flour required 8 h hydrolysis. 相似文献