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1.
S Sonnino R Ghidoni G Gazzotti G Kirschner G Galli G Tettamanti 《Journal of lipid research》1984,25(6):620-629
A semi-preparative, analytical high performance liquid chromatographic (HPLC) procedure is described for the isolation of molecular species of GM1 and GD1a gangliosides containing a single long chain base, C18 or C20 sphingosine, C18 or C20 sphinganine, each in its natural erythro or unnatural threo form. The threo forms were obtained from 2,3-dichloro-5,6-dicyanobenzoquinone/NaBH4 -treated gangliosides. The ganglioside molecular species separated by HPLC were analyzed for carbohydrate, fatty acid, and long chain base composition. In particular, long chain bases were submitted to gas-liquid chromatographic-mass spectrometric analyses as their trimethylsilyl (TMS) or N-acetyl-TMS derivatives, and chain length, presence or absence of C4-C5 double bond, and C-3 steric configuration were ascertained. The final preparations of individual molecular species of GM1 and GD1a gangliosides were more than 99% homogeneous in their saccharide moiety, contained a single long chain base (homogeneity higher than 99%), and had a fatty acid composition primarily of stearic acid (92 to 97%). All the individual molecular species of GM1 and GD1a gangliosides were also prepared in radioactive form by selective tritiation at C-3 of the long chain base. Their specific radioactivity ranged from 1.3 to 1.45 Ci/mmol. The availability of these molecular species of gangliosides is expected to facilitate studies aimed at ascertaining the role played by the hydrophobic portion in the functional behavior of gangliosides. 相似文献
2.
Preparation of GM1 ganglioside molecular species having homogeneous fatty acid and long chain base moieties 总被引:2,自引:0,他引:2
S Sonnino G Kirschner R Ghidoni D Acquotti G Tettamanti 《Journal of lipid research》1985,26(2):248-257
A new procedure is described for preparing the molecular species of GM1 ganglioside that carry a single fatty acid (myristic (C14:0), stearic (C18:0), arachidic (C20:0) or lignoceric (C24:0) acid) and a single long chain base (C18 or C20 sphingosine, C18 or C20 sphinganine, each of them in natural 3D(+)erythro or unnatural 3L(-)threo form). The procedure consisted of the following steps: a) alkaline hydrolysis of GM1 ganglioside in the presence of tetramethylammonium hydroxide, which produces de-N-acylation of the ceramide and de-N-acetylation of the sialic acid residue; b) specific re-N-acylation at the long chain base amino group with a new fatty acid (myristic, stearic, arachidic, or lignoceric) in the presence of 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride; and c) final re-N-acetylation at the level of the sialic acid residue. GM1 ganglioside molecular species, completely homogeneous in the ceramide portion, were prepared by reversed phase high performance liquid chromatography. The GM1 ganglioside molecular species were analyzed for saccharide, fatty acid, and long chain base composition by chemical and spectrometric analyses. Using a combination of the two procedures, 32 different molecular species of GM1 ganglioside, over 99% homogeneous, have been prepared. 相似文献
3.
Naoki Morita Nobuhiro Okajima Masaru Gotoh Hideyuki Hayashi Hidetoshi Okuyama Shoji Sasaki 《Archives of microbiology》1992,157(3):223-228
The activity of fatty acid synthetase (FAS) from Vibrio sp. strain ABE-1 required the presence of acyl carrier protein and was completely inhibited by thiolactomycin, an inhibitor specific for a type II FAS. These observations indicate that this enzyme is a type II FAS. Analysis by gas-liquid chromotography of the reaction products synthesized in vitro from [2-14C]malonyl-CoA by the partially purified FAS revealed, in addition to 16-and 18-carbon fatty acids which are normal constituents of this bacterium, the presence of fatty acids with very long chains. These fatty acids were identified as saturated and mono-unsaturated fatty acids with 20 up to as many as 30 carbon atoms. The longest fatty acids normally found in this bacterium contain 18-carbon atoms. These results suggest that the FAS from Vibrio sp. strain ABE-1 has potentially the ability to synthesize fatty acids with very long chains.Abbreviations ACP
acyl carrier protein
- FAME
fatty acid methyl ester
- FAS
fatty acid synthetase
- FID
flame ionization detection
- GLC
gas-liquid chromatography
- TLC
thin-layer chromatography
- In designations of fatty acids, such as 16:0, 16:1, etc
the colon separates the number that denotes the number of carbon atoms and the number that denotes the number of double bonds, respectively, in the molecule
- 16:0-CoA
CoA ester of 16:0 相似文献
4.
Alden KP Dhondt-Cordelier S McDonald KL Reape TJ Ng CK McCabe PF Leaver CJ 《Biochemical and biophysical research communications》2011,(3):51-580
Sphingolipids are ubiquitous components of eukaryotic cells and sphingolipid metabolites, such as the long chain base phosphate (LCB-P), sphingosine 1 phosphate (S1P) and ceramide (Cer) are important regulators of apoptosis in animal cells. This study evaluated the role of LCB-Ps in regulating apoptotic-like programmed cell death (AL-PCD) in plant cells using commercially available S1P as a tool. Arabidopsis cell cultures were exposed to a diverse array of cell death-inducing treatments (including Cer) in the presence of S1P. Rates of AL-PCD and cell survival were recorded using vital stains and morphological markers of AL-PCD. Internal LCB-P levels were altered in suspension cultured cells using inhibitors of sphingosine kinase and changes in rates of death in response to heat stress were evaluated. S1P reduced AL-PCD and promoted cell survival in cells subjected to a range of stresses. Treatments with inhibitors of sphingosine kinase lowered the temperature which induced maximal AL-PCD in cell cultures. The data supports the existence of a sphingolipid rheostat involved in controlling cell fate in Arabidopsis cells and that sphingolipid regulation of cell death may be a shared feature of both animal apoptosis and plant AL-PCD. 相似文献
5.
Deoxyribonucleic acid base composition of simonsiellaceae 总被引:1,自引:0,他引:1
Daisy A. Kuhn David A. Gregory Michael D. Nyby Manley Mandel 《Archives of microbiology》1977,113(3):205-207
The molar percentages of guanine plus cytosine in the DNA of 51 strains of Simonsiellaceae were determined by buoyant density ultracentrifugation of cell lysates in CsCl. The DNA base ratios ranged from 41–55 mole-% guanine plus cytosine. These values fall within the range known for the Order Cytophagales, the non-fruiting gliding bacteria, and are out-side the range of the Order Myxobacterales, the fruiting myxobacteria. Among the strains of the genus Simonsiella, four distinct groups can be delineated on the basis of source of origin (sheep, dog, cat, human) and GC content. The neotype of Alysiella filiformis has a GC content of 45.4 mole-%. 相似文献
6.
Nina V Prokazova Irina A Mikhailenko Sergey N Preobrazhensky Vadim O Ivanov Sergey N Pokrovsky Natalia G Timofeeva Maria A Martinova Vadim S Repin Lev D Bergelson 《Glycoconjugate journal》1986,3(3):273-286
The role of gangliosides in the reception of low density lipoproteins (LDL) was studied using as targets mouse ascites hepatoma 22a (MAH) cells which bind LDL through a specific high affinity receptor. Low density lipoprotein binding and uptake by MAH cells decreased after brief treatment of the cells with neuraminidase to partially remove surface sialic acid residues. The LDL uptake capability of the neuraminidasetreated MAH cells was fully restored after incorporation of exogeneous GM1- and GD1a-gangliosides into the cell surface. In contrast, free (extracellular) gangliosides inhibited LDL uptake by native MAH cells. This inhibitory effect was seen at ganglioside concentrations corresponding to the ganglioside content of serum and was most pronounced with gangliosides whose sialic acids were linked to a terminal galactose residue (GM3, GD1a, GT1b) but was smaller or absent with gangliosides whose sialic acids were attached to an internal galactose (GM1, GM2). The binding of gangliosides to LDL was structure and concentration dependent, saturable and trypsin sensitive. The LDL-ganglioside interaction was further investigated by steady state fluorescence spectroscopy. Changes in the LDL fluorescence polarization were observed with as little as 0.01 M concentrations of the gangliosides. The magnitude and nature of the effect depended on the type of ganglioside. We conclude that the LDL surface possesses sites recognizing specific carbohydrate sequences of glycoconjugates and that changes in the cell surface concentrations of sialic acids significantly modulate the LDL uptake. It is postulated that shedding of gangliosides into the blood stream may be a factor involved in regulation of cholesterol homeostasis.Abbreviations MAH
mouse ascites hepatoma 22a
- LDL
low density lipoprotein
- ASM
anthrylvinyl-labeled sphingomyelin [N-12-(9-anthryl-trans-dodecanoyl-sphingosine-1-phosphocholine]
- RITC
rhodamine isothiocyanate. The designation of gangliosides follows the IUPAC-IUB recommendation [1]: GM3, II3NeuAc-LacCer, II3-N-acetylneuraminosyllactosylceramide
- GM2
II3-NeuAc-GgOse3Cer, II3-N-acetylneuraminosylgangliotriaosylceramide
- GM1
II3-NeuAc-GgOse4Cer, II3-N-acetylneuraminosylgangliotetraosylceramide
- GD1a, II3
IV3(NeuAc)2-GgOse4Cer, II3, IV3-di(N-acetylneuraminosyl)gangliotetraosylceramide
- GT1b
II3(NeuAc)2, IV3 NeuAc-GgOse4Cer, II3-di-N-acetylneuraminosyl, IV3-N-acetylneuraminosylgangliotetraosylceramide 相似文献
7.
The DNA base compositions of 60 strains ofScenedesmus were determined and found to be a valuable indicator for the differentiation within this genus (except for the very closely related species of the subsect.Desmodesmus). The range for allScenedesmus species was 49.9–69.3 mol% GC for nuclear DNA and 36.8–39.9 mol% GC for chloroplast DNA. The separation of the genusTetradesmus cannot be verified by GC values, becauseS. obliquus andS. (Tetradesmus)wisconsinensis have a similar GC content.S. (Chlorella)ultrasquamata, S. costato-granulatus (sect.Costato-granulati) andS. lunatus are separated clearly from all other species of the genus because of their high GC content. 相似文献
8.
该研究以樟树转录组数据为基础,筛选克隆了拟南芥AtLACS9同源候选基因CcLACS9,二者序列相似性为75%。相关软件预测CcLACS9享有植物LACS亚家族成员3个特征motifs,且N端含有定位质体信手肽。在Δlacs缺陷型酵母互补测试中,以油酸作为唯一外源脂肪酸、转化了CcLACS9的突变型酵母恢复正常生长,证明CcLACS9具有典型的脂肪酰基CoA合成酶的功能。为探究CcLACS9是否参与了樟树籽油生物合成,进一步研究了其组织表达模式和在种子发育过程中其表达量与籽油累积量之间的关系。实时荧光定量PCR分析显示CcLACS9基因在种仁与花中优势表达,种仁中相对表达量是根中的17.74倍。随机测定了30棵成年樟树成熟期种子千粒重、籽油含量和中链脂肪酸比例等指标。根据仁油含量将测试群体划为高、中、低三个不同品级,并在各品级中挑选3棵单株、逐月关联分析其仁油含量与CcLACS9相对表达量。结果表明:在种仁发育前期,仁油含量和CcLACS9表达量都持续上升且二者呈正相关性,8月份为CcLACS9表达量峰值期; 9月下旬后,仁油含量趋向稳定但CcLACS9表达量仍处于较高水平但呈现下降趋势,二者无明显相关性。LACS亚家族在植物进化中较为保守,同源基因在不同植物中具有相同或相似的功能。该研究结果暗示CcLACS9可能拥有AtLACS9相似的生物学功能,即在樟树种仁油酯合成和累积过程中起重要作用。 相似文献
9.
Synthesis of medium and long chain triacylglycerols (MLCTs) by lipase-catalyzed interesterification of soybean oil with medium chain triacylglycerols (MCTs) in a solvent-free system is investigated in this study. Firstly, reaction conditions of the interesterification were optimized. Under the optimum conditions (Lipozyme 435 of 8% as catalyst, substrate ratio of 40:60, heating temperature of 90 °C, reaction time of 6 h), MLCTs were obtained in a 74.9% yield Secondly, a two-step purification was conducted to reduce acid, peroxide values and diacylglycerol content to acceptable levels. After the deacidification by the neutralization and silica gel absorption, acid, peroxide values and diacylglycerol content were reduced to 0.14 mg/g, 1.5 mmol/kg and 1.2%, respectively. The determined levels of physicochemical parameters of purified MLCTs fell within the permitted ranges of the Chinese national standard specification for MLCTs. After a two-step purification, a qualified product was produced and can be used in pharmaceutical industry as a component of fat emulsion injection. The process for the preparation of MLCTs is scalable. Differently from previous studies, the aim of this study is to obtain MLCTs products meeting the national standard JX20070100 for structured triacylglycerols. 相似文献
10.
The effect of ciprofibrate, a hypolipidemic drug, was examined in the metabolism of palmitic (C16:0) and lignoceric (C24:0) acids in rat liver. Ciprofibrate is a peroxisomal proliferating drug which increases the number of peroxisomes. The palmitoyl-CoA ligase activity in peroxisomes, mitochondria and microsomes from ciprofibrate treated liver was 3.2, 1.9 and 1.5-fold higher respectively and the activity for oxidation of palmitic acid in peroxisomes and mitochondria was 8.5 and 2.3-fold higher respectively. Similarly, ciprofibrate had a higher effect on the metabolism of lignoceric acid. Treatment with ciprofibrate increased lignoceroyl-CoA ligase activity in peroxisomes, mitochondria and microsomes by 5.3, 3.3 and 2.3-fold respectively and that of oxidation of lignoceric acid was increased in peroxisomes and mitochondria by 13.4 and 2.3-fold respectively. The peroxisomal rates of oxidation of palmitic acid (8.5-fold) and lignoceric acid (13.4-fold) were increased to a different degree by ciprofibrate treatment. This differential effect of ciprofibrate suggests that different enzymes may be responsible for the oxidation of fatty acids of different chain length, at least at one or more step(s) of the peroxisomal fatty acid -oxidation pathway. 相似文献
11.
12.
The cell walls of the yeast and mycelial forms of Yarrowia lipolytica were isolated and purified. Electron microscopy studies showed no differences between both types of cell walls. Chemical analysis revealed that the yeast cell wall contained 70% neutral carbohydrate, 7% amino sugars, 15% protein, 5% lipids and 0.8% phosphorus. Mycelial cell walls contained 70% carbohydrate, 14% aminosugars, 6% protein, 5% lipids and 0.6% phosphorus. Three polysaccharides: -glucan, mannan and chitin were detected. Proteins were solubilized from both cell wall fractions and separated by polyacrylamide gel electrophoresis. About 50 protein bands were detected, four of them corresponding to glycoproteins. The cell walls of the yeast and mycelial forms of Y. lipolytica were qualitatively similar and only quantitative differences were found.Abbreviations GlcNAc
N-acetylglucosamine
- FITC-WGA
fluorescein isothiocyanate-wheat germ agglutinin
- PAS
periodic acid Schiff 相似文献
13.
Pavlova lutheri, a marine microalga, is rich in the very long chain polyunsaturated fatty acids (VLCPUFAs) eicosapentaenoic (20:5n-3) and docosahexaenoic (22:6n-3) acids. Using an expressed sequence tag approach, we isolated a cDNA designated Pldes1, and encoding an amino acid sequence showing high similarity with polyunsaturated fatty acid front-end desaturases. Heterologous expression in yeast demonstrated that PlDES1 desaturated 22:5n-3 and 22:4n-6 into 22:6n-3 and 22:5n-6 respectively, and was equally active on both substrates. Thus, PlDES1 is a novel VLCPUFA Delta4-desaturase. Pldes1 expression is four-fold higher during the mid-exponential phase of growth compared to late exponential and stationary phases. 相似文献
14.
Manuela Valsecchi Vanna Chigorno Sandro Sonnino Guido Tettamanti 《Chemistry and physics of lipids》1992,60(3):247-252
A study has been made of the association properties of the two GM1 ganglioside molecular species GM1-C18 and GM1-C20 (containing C18 and C20 long chain bases, respectively) to rat cerebellar granule cells in culture. Both gangliosides recognized, to the same extent, and associated with them to give a form of association, the trypsin-labile form. This form was removed by treatment with trypsin enzyme. Both gangliosides associated stably with the cells to become components of the cell membranes. Although similar amounts of the two gangliosides entered the cells, being then metabolized, the time course of the association was different for the two gangliosides: after 15 h of ganglioside-cell incubation the amount of GM1-C18 inserted into the cell membrane was 2.43 times higher than that of GM1-C20. 相似文献
15.
Two forms of sucrose-phosphate synthase (EC 2.4.1.14) were resolved from leaves of three species, maize (Zea mays L. cv. Pioneer 3184), soybean (Glycine max (L.) Merr., cv. Ransom) and spinach (Spinacia oleracea L. cv. Resistoflay) by hydroxyapatite Ultrogel chromatography, using a 75-mM (designated peak 1) and 250-mM (peak 2) K-phosphate discontinuous-gradient elution. Rechromatography of the two forms showed that they were not readily interconvertible. The distribution of activity between the two forms differed among species and changed during purification of the enzyme. Recovery of peak-1 activity was specifically lowered when maize leaf extracts were prepared in the absence of magnesium, indicating that the two forms may differ in stability. In addition, the forms of the enzyme from maize differed in the extent of glucose-6-phosphate activation. These results provide evidence for the existence of multiple forms of sucrose-phosphate synthase in leaves of different species and that the forms differ in regulatory properties.Abbreviations Fru6P
fructose 6-phosphate
- Glc6P
glucose 6-phosphate
- HAU
hydroxyapatite Ultrogel
- Pi
inorganic phosphate
- SPS
sucrose-phosphate synthase
- UDP
uridine 5-diphosphate
- UDPG
uridinediphosphate glucose
Cooperative investigations of the United States Department of Agriculture, Agricultural Research Service, and the North Carolina Agricultural Research Service, Raleigh. Paper No. 10511 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh. Supported in part by USDA Competitive Research Grant No. 85-CRCR-1-1568 相似文献
16.
Vanna Chigorno Marina Pitto Gloria Cardace Domenico Acquotti Günther Kirschner Sandro Sonnino Riccardo Ghidoni Guido Tettamanti 《Glycoconjugate journal》1985,2(3-4):279-291
The preparation of a GM1-ganglioside (GM1) [14C]-labelled in the sialic acid residue is reported. This can be obtained by re-N-acetylation in the presence of [1-14C]-acetic anhydride, of a GM1 derivative de-N-acetylated specifically on the sialic acid residue by alkaline hydrolysis of GM1 with tetramethylammonium hydroxide. The radiolabelled GM1 is utilized to investigate the binding properties and the mode of interaction of GM1 with cultured fibroblasts. Three different forms of association (one serum-removable, one trypsin-removable and one trypsin-stable) have been recognized to occur in a way that depended on cell culture conditions (presence or absence of fetal calf serum), ganglioside concentration (from, 5×10–9 M to 10–4 M) and incubation time (up to 24 h). Some metabolic modifications of GM1 during the period of high cell viability were also investigated.Abbreviations GM1
GM1-ganglioside, II3NeuAc-GgOse4Cer
- FCS
fetal calf serum
- EMEM
Eaglés Minimum Essential Medium with Earlés salts
- PBS
Dulbecco phosphate buffered saline without calcium and magnesium 相似文献
17.
The sphingoid long-chain base (LCB) composition of glucosylceramides was characterized in 31 species of Fabaceae including
the model legumes Lotus japonicus and Medicago truncatula. With the exception of Lupinus texensis L, the 8-trans/cis-unsaturated isomers of 4-hydroxy-8-sphingenines [i.e., t18:1 (8t) plus t18:1 (8c)] were the major components in each species.
In tribe Fabeae, each species from four genera—Pisum, Lathyrus, Lens, and Vicia—showed that more than 50% of dihydroxy sphingoid LCBs are 8-sphingenines [i.e., d18:1 (8t) plus d18:1 (8c)]. These results
suggest that the sphingoid LCB composition of glucosylceramides reflects the phylogenetic relationships within the Fabeae. 相似文献
18.
Heterotrophic production of long chain omega-3 fatty acids utilizing algae and algae-like microorganisms 总被引:13,自引:0,他引:13
Although some interest in growing microalgae heterotrophically for the production of pigments was generated in the 1960s, only minimal commercial research was focused on this type of production technology until the 1980s. Recent developments indicating the nutritional and pharmaceutical importance of long chain omega-3 polyunsaturated fatty acids in the human diet have stimulated interest in microalgae as a source of these vital compounds, for they are the primary producers of these fatty acids in marine food webs. Food and pharmaceutical quality production can be enhanced both by the degree of process control and by the sterility achieved through a fermentation process, when compared to outdoor solar pond production. The data presented illustrate that microalgal-based heterotrophic production systems can exhibit omega-3 fatty acid productivities 2–3 orders of magnitude greater than those of outdoor pond systems. Additionally, long chain omega-3 fatty acid productivities reported for the microalgal fermentation systems are 1–2 orders of magnitude greater than productivities reported for fungal or bacterial systems. 相似文献
19.
Li J Sheng Y Tang PZ Tsai-Morris CH Dufau ML 《The Journal of steroid biochemistry and molecular biology》2006,98(4-5):207-217
Gonadotropin-regulated long chain acyl-CoA synthetase (GR-LACS) is a novel hormonally regulated fatty acyl-CoA synthetase (FACS) with activity for long-chain fatty acids. The presence of this enzyme in the Leydig cells of the mature rat testis and its mode of regulation suggest that it participates in testicular steroidogenesis. This study demonstrates that GR-LACS expression is tissue, cell and species-specific. The 79 kDa GR-LACS protein is expressed in rodent gonads and brain, and only in the mouse in the adrenal cortex. In the ovary of both species it is associated with follicles undergoing atresia. It is present in the newborn and immature testis tubules and after puberty only in the Leydig cells. A distinct GR-LACS protein species of 64 kDa that was more abundant than the 79 kDa long form was found in the rat brain. Also, a minor 73 kDa form was observed in the rat brain and mouse ovary. Two novel species resulting from alternatively splicing of the GR-LACS gene were identified in a rat brain cDNA library: a short form 1 (S1) lacking exon 8 and short form 2 (S2) lacking exons 6–8. Expression studies revealed that the sizes of the S1/S2 proteins are comparable to those of the endogenous variant species. Neither S form contains FACSs activity, suggesting that exon 8 is essential for the enzymatic function. GR-LACS variants exhibit small but significant dominant negative effects on the FACS activity of the long form. GR-LACS variants may regulate the long form's activity in the brain. 相似文献
20.
Estimates of nuclear DNA base composition by determination of thermal denaturation temperatures (Tm) indicate guanine + cytosine (G + C) levels of 35.4–46.8% for ten species of the Gracilariaceae, representing the generaGracilaria andHydropuntia. Tm values were found to be reproducible with variation among most samples and replicates of less than 1 °C and 2 mol%. Interspecific variation in G + C values was less than 11.4% amongGracilaria species. Calculation of intragenomic base pair composition distribution based on mid-resolution thermal denaturation (A 1 °C/min with 4s interval H and dT logging) indicated an inverse relationship between maximum similarity values and taxonomic rank. Intraspecific (population level) maximum similarity (homology) values were estimated to range from 79–90% inGracilaria tikvahiae (4 isolates). Interspecific values of 46–69% were found in 13 species ofGracilaria. Nucleotide distribution similarity values for the Gracilariaceae are compared with previous information for genome organization and complexity, genome size and karyotype patterns.Author for correspondence 相似文献