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21.
Effect of Phospholipase Digestion and Lysophosphatidylcholine on Dopamine Receptor Binding 总被引:4,自引:4,他引:0
[3H]Spiperone specific binding by microsomal membranes isolated from sheep caudate nucleus is decreased by trypsin and phospholipase A2 (Vipera russeli), but is insensitive to neuraminidase. The inhibitory effect of phospholipase A2 is correlated with phospholipid hydrolysis. After 15 min of phospholipase (5 micrograms/mg protein) treatment, a maximal effect is observed; the maximal lipid hydrolysis is about 56% and produces 82% reduction in [3H]spiperone binding. Equilibrium binding studies in nontreated and treated membranes showed a reduction in Bmax from a value of 388 +/- 9.2 fmol/mg protein before phospholipase treatment to a value of 52 +/- 7.8 fmol/mg protein after treatment, but no change in affinity (KD = 0.24 +/- 0.042 nM) was observed. Albumin washing of treated membranes removes 47% of lysophosphatidylcholine produced by phospholipid hydrolysis without recovering [3H]spiperone binding activity. However, the presence of 2.5% albumin during phospholipase A2 action (1.5 micrograms/mg protein) prevents the inhibitory effect of phospholipase on [3H]spiperone binding to the membranes, although 28% of the total membrane phospholipid is hydrolysed. Lysophosphatidylcholine, a product of phospholipid hydrolysis, mimics the phospholipase A2 effect on receptor activity, but the [3H]spiperone binding inhibition can be reversed by washing with 2.5% defatted serum albumin. Addition of microsomal lipids to microsomal membranes pretreated with phospholipase does not restore [3H]spiperone stereospecific binding. It is concluded that the phospholipase-mediated inhibition of [3H]spiperone binding activity results not only from hydrolysis of membrane phospholipids, but also from an alteration of the lipid environment by the end products of phospholipid hydrolysis. 相似文献
22.
Qualitative analyses were made from whole-body cuticular extracts of Scapteriscus abbreviatus, Scapteriscus acletus, Scapteriscus vicinus, and Neocurtilla hexadactyla by isothermal and temperature-programmed gas chromatography. Adults of both sexes and nymphs of each species were collected in Florida. The chromatographic profiles of peaks were distinct and easily recognizable for each species, regardless of sex or developmental stage. Distinct sexual differences were found in S. acletus and S. abbreviatus. Specimens of S. abbreviatus from Puerto Rico and S. vicinus from Bolivia produced gas chromatography (GC) traces very similar to those of conspecifics collected in Florida. Evidence is presented to illustrate the potential importance of volatile cuticular lipid analysis as a tool for mole cricket identification. Cuticular extracts of an undescribed short-winged species of Scapteriscus from Bolivia were also examined and produced GC traces unlike those of any other species analyzed to date. 相似文献
23.
A rapid one-step purification of NADPH-cytochrome c (P-450) reductase from rat liver microsomes 总被引:1,自引:0,他引:1
NADPH-cytochrome c (P-450) reductase from liver microsomes of phenobarbital-treated rats has been purified in a single step by affinity chromatography on agarose-hexane-adenosine 2',5'-diphosphate. As determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, enzyme assay, and radioimmunoassay the protein obtained by this single step procedure is as pure as that isolated by multicolumn procedures. 相似文献
24.
Abstract: Primary cultures were prepared from newborn rat brain. After 16-18 days, they consisted mainly of mature and immature astrocytes and oligodendrocytes, as judged by immunohistochemistry. To study the metabolism of ethanolamine glycerophospholipids, the cells were incubated with 1-[1-3 H]alkyl- sn -glycero-3-phosphoethanolamine (1-alkyl-GPE), for 1–20 h. Five main products were formed: 1-alkyl-2-acyl-GPE; 1-alkyl-2-acyksn-glycero-3-phosphocholine (1-alkyl-2-acyl-GPC); 1-alkenyl-2-acyl-GPE (ethanolamine plasmalogen); 1-alkenyl-2-acyl-GPC (choline plasmalogen); and 1-alkyl-glycerol. Acylation of the substrate was the main reaction during the first 3 h of incubation, whereas desaturation to plasmaiogen reached a maximum after 12 h. Greater amounts of radioactivity were observed in the phosphatidylcholine fraction after longer incubation times. Only small amounts of choline plasmalogen were observed. The phosphatidylethanolamine fraction consisted of 26.5% diacyl-, 27.5% alkyl-acyl-, and 46.0% alkenyl-acyl- compounds, whereas the corresponding data for the phosphatidylcholine fraction were 78.5, 16.4, and 5.1%, respectively, after 20 h of incubation. Hydrolysis of the substrate to 1-alkyl-glycerol was a minor reaction. 相似文献
25.
The fatty acid, sterol and chlorophyll composition of the calcified, unicellular alga Thoracosphaera heimii (Lohmann) Kamptner are reported. The presence of 4,23,24-termethyl-5α-cholest-22E-en-3β-ol (dinosterol), 4,23,24-trimethyl-5α-cholest-22E-en-3-one (dinosterone) and the predominance of C18, C20 and C22 unsaturated fatty acids, including the acid 18:5ω3, indicates that T. heimii is a dinoflagellate. The fatty acid: sterol ratio (1.3), is typical of dinoflagellates. The geochemical significance of dinosterone, the high relative concentration of 4-desmethyl-5α-stanols and the role of 23-methyl-5α-cholest-22E-en-3β-ol in the biosynthesis of dinosterol in T. heimii are also discussed. 相似文献
26.
Analyses of the sulphatides in the pollen and style of Oenothera missouriensis show that these membranous lipids are comparatively less important in the styles than in the pollen. Incompatible pollination is followed by a large increase in sulphatides, whereas cross-pollination also causes an increase in sulphatide but to a much lesser extent. This mobilization of sulphatides in the membrane is discussed in term of permeability. 相似文献
27.
Z. Krupa 《Photosynthesis research》1983,4(3):229-239
Thylakoid membranes obtained from bean chloroplasts treated with bean galactolipase or phospholipase A2 (from Crotalus terr. terr.) showed marked changes in their polypeptide patterns when separated on SDS-PAGE. The obtained results have been discussed with regard to the relationship between chloroplast lipids and polypeptides originating from chlorophyll-protein complexes of bean thylakoids. A coexistence between galactolipids and the peripheral antennae in PS I complex and LHCP3 as well as a conspicuous role of phospholipids in PSI and PSII centre chlorophyll-protein complexes has to be underlined.Abbreviations CP1
chlorophyll a-protein complex of PSI
- CPa
chlorophyll a-protein complex of PSII
- D10
digitonin subchloroplast particles enriched in PSII
- D144
digitonin subchloroplast particles enriched in PSI
- DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- LHCP1–3
light harvesting chlorophyll a/b protein complexes
- PAGE
polyacrylamide gel electrophoresis
- PSI
photosystem I
- PSII
photosystem II
- SDS
sodium dodecyl sulphate
- TCA
trichloroacetic acid
- Tricine
N-Tris-(hydroxymethyl)-methylglycine
- Tris
Tris-(hydroxymethyl)-aminomethan 相似文献
28.
The electrical conductivity of bilayer lipid membranes (BLM) of oxidized cholesterol has been measured separately in bathing solutions of sodium sulphate, sodium chloride, sodium bromide, sodium iodide and also in bathing solutions of iodine and iodine containing these salts. An attempt has been made to explain the conduction of electric current across the membranes. 相似文献
29.
In anoxic salt marsh sediments of Sapelo Island, GA, USA, the vertical distribution of CH4 production was measured in the upper 20 cm of surface sediments in ten locations. In one section of high marsh sediments,
the concentration and oxidation of acetate in sediment porewaters and the rate and amount of14C acetate and14CO2 incorporation into cellular lipids of the microbial population were investigated. CH4 production rates ranged from <1 to 493 nM CH4 gram sediment−1 day−1 from intact subcores incubated under nitrogen. Replacement with H2 stimulated the rate of methane release up to nine fold relative to N2 incubations. Rates of lipid synthesis from CO2 averaged 39.2 ×10−2nanomoles lipid carbon cm3 sediment−1 hr−1, suggesting that CO2 may be an important carbon precursor for microbial membrane synthesis in marsh sediments under anoxic conditions. Qualitative
measurements of lipid synthesis rates from acetate were found to average 8.7 × 10−2 nanomoles. Phospholipids were the dominant lipids synthesized by both substrates in sediment cores, accounting for an average
of 76.6% of all lipid radioactivity. Small amounts of ether lipids indicative of methanogenic bacteria were observed in cores
incubated for 7 days, with similar rates of synthesis for both CO2 and acetate. The low rate of ether lipid synthesis suggests that either methanogen lipid biosynthesis is very slow or that
methanogens represent a small component of total microbial lipid synthesis in anoxic sediments.
present address: The University of Maryland,, Chesapeake Biological Laboratory, Box 38, Solomons, MD 20688, USA 相似文献
30.
Validation of signature polarlipid fatty acid biomarkers for alkane-utilizing bacteria in soils and subsurface aquifer materials 总被引:3,自引:0,他引:3
David B. Ringelberg John D. Davis Glen A. Smith Susan M. Pfiffner Peter D. Nichols Janet S. Nickels J.Michael Henson John T. Wilson Marylynn Yates Donald H. Kampbell Harvey W. Read Thomas T. Stocksdale David C. White 《FEMS microbiology letters》1989,62(1):39-50
Abstract Extractable cell membrane-derived polarlipid ester-linked fatty acids (PLFA) obtained from aerated soils gassed with methane or propane and from methane- and propane-oxidizing bacteria isolated from the soils were analyzed by capillary gas chromatography/mass spectrometry. Exposure of aerated soils to methane resulted in the formation of a high proportion of an unusual 18-carbon mono-unsaturated PLFA, 18:lw8c. High proportions of this fatty acid biomarker are found in monocultures from this soil grown in minimal media with methane. This PLFA has been previously established as associated with authentic type II methane-oxidizing bacteria. The microbiota in aerated soils exposed to hydrocarbons containing propane, formed a suite of PLFA characterized by high proportions of a 16-carbon mono-unsaturated acid, 16:lw6c, and an 18-carbon saturated fatty acid with an additional methyl branch at the 10 position, 10 Me 18:0. This PLFA pattern has been detected in several monocultures enriched from the soil with propane-amended minimal media. The correspondence of high proportions of these unusual mono-unsaturated PLFA in the isolated monocultures and in situ in the soils after stimulation with the appropriate hydrocarbon is a strong validation of the utility of these biomarkers in defining the community structure of the surface soil microbial community. 相似文献