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1.
Sarcina marina (NCMB 778) grew over the temperature range 20–45°C but no growth was recorded at 15°C or 50°C. At the optimum growth temperature of 34°C the doubling time was 14.5 h.The major polar lipid components, tentatively identified as the diether analogues of phosphatidyl glycerophosphate (PGP), phosphatidyl glycerol (PG), diglycosyl diglyceride (DGD) and triglycosyl diglyceride (TGD), and the major neutral lipid components, tentatively identified as squalene, dihydrosqualene, tetrahydrosqualene, vitamin MK8, geranyl geraniol and di-O-phytanyl glycerol, are identical to those found in other extremely halophilic rods and cocci.The total lipid content varied with growth conditions from 0.6 – 3.2% of the dry cell weight, polar lipids accounted for between 94.3 and 83.6% of the total lipid, the remainder being neutral lipid.In response to both the transition from exponential to stationary phase and a reduction of 14°C in growth temperature, batch cultures showed: (i) an increase in total lipid content; (ii) a decrease in PG and (iii) an increase in PGP. Specific responses to the temperature decrease were (i) increased total lipid content; (ii) no decrease in neutral lipids in stationary phase; (iii) marked reduction in PG and (iv) raised DGD. (i) and (ii) could be mechanisms for increasing membrane fluidity.In common with all other extreme halophiles investigated the alkyl side chains of S. marina polar lipids were identified as the phytanyl (3R, 7R, 11R, 15-tetramethylhexadecyl) group. Its structure did not appear to vary with temperature so that the normal mechanisms for modifying the structure of lipid alkyl side chains to modulate membrane fluidity in response to temperature changes probably does not occur in this group of microorganisms.  相似文献   

2.
[1-14C]-Acetate incorporation into total and polar lipids was studied in the growing pollen tubes of Crotalaria juncea. Ungerminated pollen had phosphatidyl inositol, phosphatidyl serine, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol, monogalactosyl diglyceride, digalactosyl diglyceride, sulpholipid and steryl glycosides. In the growing pollen tubes considerable [1-14C]-acetate incorporation was observed into the individual polar lipids. The exogenous carbon source significantly influenced lipid biosynthesis. Boric acid (20mg/l.) promoted both pollen tube growth and acetate incorporation into phospholipids. In comparison to 5′-adenosine monophosphate, cyclic-3′,5′-adenosine monophosphate (cAMP) promoted tube growth and also enhanced phospho-and glycolipid biosynthesis. The regulation of membrane component biosynthesis by cAMP is suggested.  相似文献   

3.
The total lipid content of Acremonium alabamensis and Thermomucor indicae-seudaticae ranged 2.6–7.3 and 8.5–13.0% of dry mycelium, respectively during development. Neutral lipid fraction increased during growth while polar and phospholipids declined. Both moulds contained palmitic, oleic, linoleic and palmitoleic acids as major fatty acid components in lipids. Degree of unsaturation of lipids of A. alabamensis was greater than that of T. indicae-seudaticae. Neutral lipids were more unsaturated than the polar lipids. The ratio of unsaturation index of polar lipids to neutral lipids was either one or less than one. The principal phospholipids of these moulds were phosphatidyl choline, phosphatidyl ethanolamine and phosphatidic acid. However, phosphatidic acid was not found in very high amounts as observed in Humicola grisea var. thermoidea.  相似文献   

4.
Conditions were defined for producing protoplasts with lysozyme and isolating the protoplast membranes from cells of Bacillus cereus T harvested late in the exponential growth phase just before sporogenesis. The membranes contained approximately 60% protein, 30% lipid, 6% carbohydrate, and 1% ribonucleic acid. Seventeen proteins were distinguished by molecular size in the membrane solubilized with sodium dodecyl sulfate, and 12 in that with phenol and acetic acid. The lipid fraction consisted of neutral lipids (28%) and phospholipids (72%). Four phospholipids were identified: diphosphatidyl glycerol, phosphatidyl ethanolamine, phosphatidyl glycerol, and lysophosphatidyl ethanolamine. Eighteen fatty acids were identified, with a predominance of branched C(15) and C(17) and of normal C(16) acids. The carbohydrate fraction consisted of neutral hexoses. A clear supernatant solution from the solubilized preparation became reaggregated into membrane by dialysis in the presence of MgCl(2). The reaggregated membrane had the same main components as the native membrane, but the amount and ratio of protein and lipid depended on the buffer and the MgCl(2) concentration. By electron microscopy, the reaggregated membranes appeared as vesicles or sheets, depending on the MgCl(2) concentration. Hexagonal lattices were occasionally detected in the negatively stained ultrastructure of both native and reaggregated membrane fragments.  相似文献   

5.
C. Kanno 《Protoplasma》1990,159(2-3):184-208
Summary The lactating mammary gland is one of the most highly differentiated and metabolically active organs in the body. Membranes of the lactating mammary cell have important roles in transmitting from one membrane to another of hormonal information and in milk secretion, which is the final event. During milk secretion, the projection of the surface membrane into the alveolar lumen by enveloping intracellular lipid droplets with the apical plasma membrane is one of the most remarkable aspects of biological membrane action throughout nature.This review focuses on current knowledge about membranes in the lactating mammary gland. (1) Advances in the isolation and properties of membranes, especially the plasma membrane and Golgi-derived secretory vesicles, concerned with milk secretion from the lactating mammary gland are described. (2) Milk serum components are secreted by fusing the membranes of secretory vesicles that condense milk secretions with the plasma membrane in the apical regions. This occurs through the formation of a tubular-shaped projection and vesicular depression in a ball-and-socket configuration, as well as by simple fusion. (3) Intracellular lipid droplets are directly extruded from the mammary epithelial cells by progressive envelopment of the plasma membranes in the apical regions. (4) The balance between the surface volume lost in enveloping lipid droplets and that provided by fusion of the secretory vesicle and other vesicles with the apical plasma membrane is discussed. (5) The membrane surrounding a milk fat globule, which is referred to as the milk fat globule membrane (MFGM), is composed of at least the coating membrane of an intracellular lipid droplet, of the apical plasma membrane and secretory vesicle membrane, and of a coat material. Consequently, MFGM is molecularly different from the plasma membrane in composition. (6) MFGM of bovine milk is structurally composed of an inner coating membrane and outer plasma membrane just after segregation. These two membranes are fused and reorganized through a process of vesiculation and fragmentation to stabilize the fat globules. Hypothetical structural models for MFGM from bovine milk fat globules just after secretion and after rearrangement are proposed.Abbrevations MFGM milk fat globule membrane - HEPES N-2-hydroxylpiperazine-N-2-ethanesulfonic acid - INT 2-(p-indophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium - SDS-PAGE polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate - Sph sphingomyelin - PC phosphatidyl choline - PE phosphatidyl ethanolamine - PS phosphatidyl serine - PI phosphatidyl inositol - PAS periodic acid-Schiff reagent - CB Coomassie brilliant blue R-250 Dedicated to Professor Stuart Patton on the occasion of his 70th birthday.  相似文献   

6.
The isolation procedure and characterization of the outer and inner membranes from Selenomonas ruminatium cells, a strictly anaerobic bacterium, are described. The metabolic fate of [14C]decanoate incorporated into the outer and inner membranes was examined. The percent distribution of radioactivities in the outer and inner membranes was about 40 and 50% of the total incorporated activity, respectively. Approximately 47% of the radioactivity incorporated into the outer membrane was recovered in the phospholipid fraction, and the remaining radioactivity was found in both aqueous and phenol layers when the outer membrane was treated with phenol-water. In contrast to [14C]decanoate, the percent distribution of [3H]glycerol in the outer and inner membranes was about 25 and 70% of the total incorporated activity, respectively. Most of the assimilated 3H was located in the phospholipid fraction of both membranes. However, no significant label was detected in either the protein or cell wall fraction. The following observations were made concerning lipid compositions in the outer and inner membranes by chemical and isotopic analyses. (i) The outer and inner membranes contained no detectable phosphatidyl glycerol or cardiolipin. (ii) A prominent radioactive compound, designated band III lipid, was found mainly in the outer membrane as a major radioactive spot when cells were grown with [14C]decanoate. This lipid contained phosphorus, 2-keto-3-deoxyoctulosonic acid and 3-OH fatty acid but no detectable glycerol. This lipid was identified tentatively to be 2-keto-3-deoxyoctulosonic acid-lipid A. (iii) Although the ubiquity of phosphatidyl ethanolamine plasmalogen in both outer and inner membranes was confirmed, the occurrence of the molecular species of phosphatidyl ethanolamine plasmalogen was quite different in the outer and inner membranes.  相似文献   

7.
Glyoxysome, endoplasmic reticulum, mitochondria, and proplastid fractions were isolated from endosperm of castor beans (Ricinus communis) germinated for 5 days at 30 C. Samples from sucrose density gradients were diluted with 0.15 m KCI and the membranes pelleted. Lipid extracts of these membranes were analyzed for phosphoglyceride, acyl lipid, and sterol content. The endoplasmic reticulum contains 1.24 mumol of phosphoglyceride per mg of protein; the mitochondria, 0.65 mumol/mg; and the glyoxysome membranes, 0.55 mumol/mg. Phosphatidyl choline and phosphatidyl ethanolamine are the most abundant lipids in all membranes studied, accounting for 70% or more of the lipid phosphorus and 50% or more of the fatty acid. Glyoxysome membranes and endoplasmic reticulum also contain phosphatidyl inositol (respectively, 9 and 17% of the lipid phosphorus) and free fatty acids (13% of the total fatty acid in each). Compared with other organelles, mitochondrial membranes have more phosphatidyl ethanolamine relative to phosphatidyl choline and are characterized by the presence of cardiolipin, in which 80% of the fatty acid is linoleate. The relative amounts of linoleate, palmitate, oleate, stearate, and linolenate in each of the phosphotoglycerides are constant regardless of the membrane source. Stimasgasterol and beta-sitosterol are present in the membranes (1-9 nmol each/mg protein).The data provide further evidence that glyoxysome membranes are derived from the endoplasmic reticulum but at the same time indicate some differentiation.  相似文献   

8.
Summary The lipid content and composition from an axolemma-rich preparation isolated from squid retinal axons was analyzed.The lipids, which accounted for 45.5% of the dry weight of this membrane, were composed of 22% cholesterol, 66.7% phospholipids and 5.2% free fatty acids. The negatively charged species phosphatidyl ethanolamine (37%), phosphatidyl serine (10%) and lysophosphatidyl ethanolamine (4%) made up 51% of the phospholipids. The amphoteric phosphatidyl choline and sphingomyelin accounted for 39% and 4%, respectively.The relative distribution of fatty acids in each of the isolated phospholipids was studied. The most remarkable feature of these phospholipids was the large proportion of long-chain polyunsaturated fatty acids. The 226 acyl chain accounted for 37% in phosphatidyl ethanolamine, 21.7% in phosphatidyl choline, 17.5% on phosphatidyl serine and 20.3% in sphingomyelin (all expressed as area %).The molar fraction of unsaturated fatty acids reached 65% in phosphatidyl ethanolamine and 42.0 and 44.8% in phosphatidyl choline and phosphatidyl serine, respectively. The double bond index in these species varied between 1.0 and 2.6.The lipid composition of the axolemma-rich preparation isolated from squid retinal axons appears to be similar to other excitable plasma membranes in two important features: (a) a low cholesterol/phospholipid molar ratio of 0.61; and (b) the polyunsaturated nature of the fatty acid of their phospholipids.This particular chemical composition may contribute a great deal to the molecular unstability of excitable membranes.The preceding papers of this series were published inArchives of Biochemistry and Biophysics.  相似文献   

9.
Perturbations induced by a toxic lectin (ricin) on lipid organisation of model membranes prepared with DPPC and DPPC-cerebrosides mixtures have been analysed by Raman and infrared spectroscopy, two powerful and non-invasive methods. Our approach involves the observation of changes in the vibrational spectra of liquid multilayers in the PO 2 - , C=0 and CH2 spectral regions for two lipid: ricin molar ratios (225:1, 75:1).The interfacial and polar regions of the multilayers, analysed by FTIR, appear to be perturbed by the protein. With both kinds of membranes, ricin mainly perturbs the C=0 ester groups of the sn-2 acylchain of DPPC. In the PO 2 - stretching region, the frequency shifts are correlated with changes in polar group hydration.In the hydrophobic core of the multilayer membrane studied by Raman spectroscopy, the interaction of ricin is associated with changes in lipid packing. These perturbations depend upon the lipid composition of the membrane. With DPPC membranes, an affect is detected at temperatures lower than T m .It corresponds to a decrease of the lipid ordering. With DPPC-cer membranes, the protein increases the acylchain packing order regardless of the temperature of the experiments (10°C<T<75°C). No perturbation of T m is observed after addition of ricin to either DPPC or DPPC-cer membranes.The different perturbations detected by Raman and FTIR suggest that ricin mainly interacts with the interfacial domains of the membranes.  相似文献   

10.
The distribution of phospholipids in the membranes of Mycobacterium phlei has been studied by the use of phospholipase C and trinitrobenzenesulfonic acid. In inverted membrane vesicles, whose external surface apparently corresponds topologically to the cytoplasmic surface of the membrane in intact cells, 80% of the phosphatidyl ethanolamine, 24% of diphosphatidyl glycerol, and 13% of phosphatidyl inositol are accessible to cleavage by phospholipase C. These results are in agreement with the finding that 70–75% of phosphatidyl ethanolamine in the membrane is accessible to chemical modification by trinitrobenzenesulfonic acid or dimethylsuberimidate at 4 °C. It can be inferred that in the inverted membrane the majority of phosphatidyl ethanolamine is present on the outer half of the lipid bilayer while inner half constitutes primarily other phospholipids namely phosphatidyl inositol and diphosphatidyl glycerol. Phospholipase C treatment of ETP membranes selectively impairs the active transport of Ca2+ without affecting the generation of a proton gradient, respiration, and coupled phosphorylation.  相似文献   

11.
The effect of adaptation to saline growth of a fresh water cyanobacterium Synechococcus 6311 on components of the cytoplasmic membranes and thylakoids was investigated. Significant changes in membrane surface charge, lipid, fatty acid, and carotenoid composition were observed upon transfer of the cells from a low salt (0.015 M NaCl) to a high salt (0.50 M NaCl) growth medium. Very similar changes in the polar lipid classes and fatty acid composition were observed in both membranes, but changes in fluidity and surface charge and a significant shift in the protein to lipid ratio were only apparent in the cytoplasmic membranes. The fluidity and surface charge data correlate well with functional studies and we can attribute the cytoplasmic membrane as the major site of interaction and adaptation to the saline environment.  相似文献   

12.
A cholestane spin probe was used to study the effects of the all-trans isomers of retinal and retinol, and the 13-cis- isomer of retinal on the degree of organization of oriented phospholipid multibilayer model membranes in the dark. Concentration-dependent effects were observed indicative of changes in the degree of organization of phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine, sphingomyelin, and brain lipids. In general the all-trans compounds improved the degree of order of the lipid films at low concentrations, and decreased it at high concentrations. The 13-cis retinal served only to decrease the degree of order. The magnitudes of these effects depend on lipid composition and the nature of the pigment polar residue. Chemical as well as nonbonding interactions are considered to be responsible for these various effects.  相似文献   

13.
Dehydrins are intrinsically disordered plant proteins whose expression is upregulated under conditions of desiccation and cold stress. Their molecular function in ensuring plant survival is not yet known, but several studies suggest their involvement in membrane stabilization. The dehydrins are characterized by a broad repertoire of conserved and repetitive sequences, out of which the archetypical K-segment has been implicated in membrane binding. To elucidate the molecular mechanism of these K-segments, we examined the interaction between lipid membranes and a dehydrin with a basic functional sequence composition: Lti30, comprising only K-segments. Our results show that Lti30 interacts electrostatically with vesicles of both zwitterionic (phosphatidyl choline) and negatively charged phospholipids (phosphatidyl glycerol, phosphatidyl serine, and phosphatidic acid) with a stronger binding to membranes with high negative surface potential. The membrane interaction lowers the temperature of the main lipid phase transition, consistent with Lti30's proposed role in cold tolerance. Moreover, the membrane binding promotes the assembly of lipid vesicles into large and easily distinguishable aggregates. Using these aggregates as binding markers, we identify three factors that regulate the lipid interaction of Lti30 in vitro: (1) a pH dependent His on/off switch, (2) phosphorylation by protein kinase C, and (3) reversal of membrane binding by proteolytic digest.  相似文献   

14.
We designed an experimental approach to differentiate the kinetics of protein binding to a lipid membrane from the kinetics of the associated conformational change in the protein. We measured the fluorescence intensity of the single Trp6 in chicken liver bile acid-binding protein (L-BABP) as a function of time after mixing the protein with lipid membranes. We mixed the protein with pure lipid membranes, with lipid membranes in the presence of a soluble quencher, and with lipid membranes containing a fluorescence quencher attached to the lipid polar head group. We fitted simultaneously the experimental curves to a three-state kinetic model. We conclude that in a first step, the binding of L-BABP to the interfacial region of the anionic lipid polar head groups occurred simultaneously with a conformational change to the partly unfolded state. In a second slower step, Trp6 buried within the polar head group region, releasing contacts with the aqueous phase.  相似文献   

15.
SYNOPSIS Fatty acids were examined of photosynthetic and non-photosynthetic marine and freshwater cryptomonads cultured as photoauxotrophs, photoheterotrophs and heterotrophs at various incubation temperatures and constant light intensity. Photo-synthetic marine and freshwater forms contained octadecatrienoic, octadecatetraenoic, eicosapentaenoic and docosahexaenoic (all-cis, ω3 acids) as the major polyunsaturates, and a freshwater heterotroph contained mostly the octadecatrienoic acid. The polar lipids of a marine, photosynthetic form, Cryptomonas sp., included the usual thylakoid membrane lipids of the chloroplasts of eukaryotic, photosynthetic cells: galactosyl diglycerides, phosphatidyl glycerol and a sulfolipid. Also present were 2 choline-containing phospholipids: phosphatidyl choline and an unknown. Ninhydrin-positive and inositol-containing lipids were not detected. Octadecatetraenoic acid comprised 75% of the total fatty acids of the monogalactosyl diglyceride fraction. The phosphatidyl glycerol was acylated mostly by ω13 trans-hexadecaenoic acid and the eicosapentaenoic acid. Evolutionary relationships of cryptomonads as mirrored in lipid composition are discussed.  相似文献   

16.
When the major polar lipid of purple membrane, a dialkyl analogue of phosphatidyl glycerophosphate, is treated with phospholipase D under the usual assay conditions for this enzyme, the reaction yields dialkylglycerol and glycerol bisphosphate, i.e. the kind of products that would be expected from a phospholipase C reaction. The effect is seen both in native purple membranes and with the pure phospholipid in the form of liposomes. The specific activity and kinetic parameters Km and Vmax of phospholipase D for the purple membrane phospholipid are similar to those for egg phosphatidylcholine. The presence of phospholipase C impurities in the phospholipase D preparations has been ruled out as an explanation for the above observations. A hypothesis is suggested, taking into account the peculiar headgroup structure of the bacterial lipid, to explain the seemingly anomalous enzyme behavior.  相似文献   

17.
In the previous paper (Block, M. A., Dorne, A.-J., Joyard, J., and Douce, R. (1983) J. Biol. Chem. 258, 13273-13280), we have described a method for the separation of membrane fractions enriched in outer and inner envelope membranes from spinach chloroplasts. The two envelope membranes have a different weight ratio of acyl lipid to protein (2.5-3 for the outer envelope membrane and 0.8-1 for the inner envelope membrane). The two membranes also differ in their polar lipid composition. However, in order to prevent the functioning of the galactolipid:galactolipid galactosyltransferase during the course of envelope membrane separation, we have analyzed the polar lipid composition of each envelope membrane after thermolysin treatment of the intact chloroplasts. The outer envelope membrane is characterized by the presence of high amounts of phosphatidylcholine and digalactosyldiacylglycerol whereas the inner envelope membrane has a polar lipid composition almost identical with that of the thykaloids. No phosphatidylethanolamine or cardiolipin could be detected in either envelope membranes, thus demonstrating that the envelope membranes, and especially the outer membrane, do not resemble extrachloroplastic membranes. No striking differences were found in the fatty acid composition of the polar lipids from either the outer or the inner envelope membrane. The two envelope membranes also differ in their carotenoid composition. Among the different enzymatic activities associated with the chloroplast envelope, we have shown that the Mg2+-dependent ATPase, the UDP-Gal:diacylglycerol galactosyltransferase, the phosphatidic acid phosphatase, and the acyl-CoA thioesterase are associated with the inner envelope from spinach chloroplasts whereas the acyl-CoA synthetase is located on the outer envelope membrane.  相似文献   

18.
A recently proposed model for proton leakage across biological membranes [Prog. Lipid Res. 40 (2001) 299] suggested that hydrocarbons specifically in the center of the lipid bilayer inhibit proton leaks. Since cellular membranes maintain a proton electrochemical gradient as a principal energy transducer, proton leakage unproductively consumes cellular energy. Hydrocarbons in the bilayer are widespread in membranes that sustain such gradients. The alkaliphiles are unique in that they contain up to 40 mol% isoprenes in their membranes including 10-11 mol% squalene [J. Bacteriol. 168 (1986) 334]. Squalene is a polyisoprene hydrocarbon without polar groups. Localizing hydrocarbons in lipid bilayers has not been trivial. A myriad of physical methods including fluorescence spectroscopy, electron-spin resonance, nuclear magnetic resonance as well as X-ray and neutron diffraction have been used to explore this question with various degrees of success and often contradictory results. Seeking unambiguous evidence for the localization of squalene in membranes or lipid bilayers, we employed neutron diffraction. We incorporated 10 mol% perdeuterated or protonated squalane, an isosteric analogue of squalene, into stacked bilayers of dioleoyl phosphatidyl choline (DOPC) doped with dioleoyl phosphatidyl glycerol (DOPG) to simulate the negative charges found on natural membranes. The neutron diffraction data clearly show that the squalane lies predominantly in the bilayer center, parallel to the plane of the membrane.  相似文献   

19.
Mesosomal vesicles and plasma membranes were isolated from Staphylococcus aureus ATCC 6538P by protoplasting and differential centrifugation. The lipids of each of the two membrane fractions were extracted with pyridine-acetic acid-N-butanol, and the nonlipid contaminants were removed by Sephadex treatment. The lipids were then separated by passage through diethylaminoethyl-cellulose columns and characterized by thin-layer chromatographic, chemical, and spectral analyses. The lipids were separated into four discrete diethylaminoethyl fractions: (i) vitamin K2, carotenoids, C55 isoprenoid alcohol, and monoglucosyl diglyceride; (ii) cardiolipin, carotenoids, phosphatidyl glycerol, diglucosyl diglyceride, and an unidentified ninhydrin-positive component; (iii) cardiolipid and phosphatidyl glyderol; (iv) cardiolipin, phosphatidyl glycerol, and phosphatidyl glucose. Qualitatively, no difference in lipid composition between mesosomal vesicles and plasma membranes was found. However, based on equal dry weights of membrane materials, a relative quantitative difference in the amount of specific lipids in mesosomal vesicles and plasma membranes was observed. There are 4 times more monoglucosyl diglyceride, 2.6 times more diglucosyl diglyceride, 3.8 times more phosphatidyl glucose, 2 times more carotenoids, and 2 times more vitamin K2 found in mesosomal vesicles than in plasma membranes. The concentration of cardiolipin and phosphatidyl glycerol is 3.6 and 6 times greater, respectively, in mesosomal vesicles.  相似文献   

20.
Abstract

Of the 14C labelled sugars (sucrose, glucose and fructose) and 14C proline, the incorporation of label into different lipid types was least from proline while sucrose was the preferred precursor over glucose or fructose. High incorporation into phosphatidyl inositol with 14C sucrose suggested that this phosphatide besides being a membrane component served possibly as the inositol storage component. High incorporation of label into phosphatidyl choline also suggested synthesis of new membranes during pollen tube growth.  相似文献   

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