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番茄叶片胞外β-N-乙酰氨基己糖苷酶的部分性质研究 总被引:1,自引:0,他引:1
用从系统感染TMV(tobacco m osaic virus)的番茄叶胞外蛋白提取液中纯化的β-N-乙酰氨基己糖苷酶为材料,研究了该酶的部分性质。以p-硝基苯基-N-乙酰-β-D-氨基葡萄糖苷(pNP-β-D-GlcNAc)或p-硝基苯基-N-乙酰-β-D-氨基半乳糖苷(pNP-β-D-GalNAc)为底物,该酶的最适pH在4.8—5.0 之间,最适温度在44—47℃之间。对酶的热稳定性研究表明,温度对酶的热变性作用表现为两相变性曲线,Km 值分别为0.36 m m ol/L (pNP-β-D-GlcNAc为底物)和0.67 m m ol/L (pNP-β-D-GalNAc 为底物),N-乙酰氨基葡萄糖(GlcNAc)和N-乙酰氨基半乳糖(GalNAc)是酶活性的竞争性抑制剂,Ag+ 和Hg2+ 是酶活性的强抑制剂,金属离子Fe2+ 、Fe3+ 和Cu2+ 也抑制酶活性。 相似文献
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为了研究Ⅱ型糖尿病(NIDDM)患者红细胞膜血型糖蛋白A(GPA)的基因表达,采用不连续聚蔗糖(Percol)密度梯度法分离15名正常人和25例NIDDM患者外周血的网织红细胞,提取其总RNA,继以Northern斑点杂交观测此两组的红细胞膜GPA基因的表达水平.采用碱性磷酸酶酶联免疫检测完成红细胞膜GP的Western印迹.结果表明,NIDDM患者组红细胞膜GPA的mRNA含量较正常人组显著增加(分别为11.92±10.5和10.18±1.08积分光密度,P<0.01).免疫印迹图谱显示,有6例NIDDM患者的区带3a、3b界限模糊,3例的区带3a、3b和4明显浅染.推测NIDDM患者红细胞膜GPA减少可能引起其基因表达呈代偿性增强,而障碍或许出现在翻译环节上,这些与NIDDM患者红细胞变形能力(RCD)的明显降低甚为相符. 相似文献
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分离纯化了人胎盘纤连蛋白(Fn),经SDS-PAGE鉴定为一条带,纯化Fn仍保持其搞原性,得率为38.7%。根据植物凝集素识别专一糖链结构的原理,应用斑点印迹法,亲和层析法和Western转移电泳研究糖链结构,结果证实:1.人胎盘Fn分子中含有复杂型N糖链(包括二天线和大于二天线的结构)以及高甘露糖型和/或杂合型N糖链;复杂型N糖链中含有平分型GlcNAc,糖链末端也可连有唾液酸;2.胰糜蛋白酶水解而获得的明胶结合片段(44kD)含有二天线和多天线复杂型糖链,也可接有平分型GlcNAc;3.肝素结合片段(30kD)以及明胶、肝素均不结合的Fn片段不含有多天线复杂型N糖链。 相似文献
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山豆根木葡聚糖的研究 总被引:5,自引:0,他引:5
研究了山豆根中一种木葡聚糖的结构.用1mol/LNaOH提取,DEAE-SephadexA-25离子交换柱层析,Fehling试剂分级得到山豆根木葡聚糖组分SSb-1FA,用完全酸水解,甲基化分析,部分酸水解,三氧化铬氧化和1H,13CNMR等方法对其结构进行研究.结果表明:SSb-1FA的分子量为2.6×104,比旋光度[α]20D=+10.9°(c0.22,H2O),由L-Fuc,D-Xyl,D-Gal和D-Glc组成,摩尔比为:2.929.97.559.8.SSb-1FA由1→4连接的β-D-Glc残基构成主链,分枝有α-D-Xyl(1→,β-D-Gal(1→2)α-D-Xyl(1→等类型,部分非还原末端由L-Fuc(1→构成. 相似文献
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禾本科植物的起源,进化及分布 总被引:6,自引:0,他引:6
禾本科植物的起源、进化及分布韩建国樊奋成李枫(中国农业大学草地研究所,北京100094)ORIGIN,EVOLUTIONANDDISTRIBUTIONOFTHEGRAMINEAEHanJian-guoFanFen-chengLiFeng(Grassl... 相似文献
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人食管鳞状上皮癌糖复合物表达的研究 总被引:1,自引:0,他引:1
凝集素可与其相对应的糖复合物特异性结合。在癌症形成过程中细胞表面经历着显著的变化,本文用十种生物素化凝集素即UEA-I、RCA-I、DBA、PSA、PNA、BSL、LCA、WGA、ConA和SBA作为探针,对正常食管上皮和食管鳞状上皮癌进行研究,以确定正常食管上皮和食管鳞状上皮癌中糖复合物的改变,结果发现,PNA和PSA在正常食管和食管鳞状上皮癌中均无反应,RCA-I、DBA、BSL、LCA、ConA和SBA受体在正常和癌组织中有着特征性变化,BSL和DBA在食管中无反应,LCA、SBA和RCA-1正常食管和食管鳞状上皮癌中反应方式不同,ConA和WGA则在同一癌组织的不同部位都显示出不同的反应。尽管UEA-I在正常食管和食管鳞状上皮癌均呈阳性反应,但似乎未表现出有意义的变化。上述结果提示食管癌的发生与含α-D-GalNAc(DBA和SBA)、β-D-Gal/β-N-acetyl-D-Galactosamine(RCA-I)、α-D-Glc/α-D-Man(LCA和ConA)、[β-(1-4)-D-GlcNAc]2/NeuNAc(WGA)和α-D-Gal(BSL)等的糖复合物的改变有较为密切的关系。 相似文献
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六例成年川金丝猴心电图的分析刘诗峰,魏武科,刘德乾,高远宏,毕洁(西北大学生物系西安710069)郭法平,范培中,雷晓青(陕西省实验林场)ANANALYSISOFELECTROCARDIOGRAMOF6ADULTRhinopithecusroxell... 相似文献
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Uemura Y Asakuma S Yon L Saito T Fukuda K Arai I Urashima T 《Comparative biochemistry and physiology. Part A, Molecular & integrative physiology》2006,145(4):468-478
Milk of an Asian elephant (Elephas maximus), collected at 11 days post partum, contained 91 g/L of hexose and 3 g/L of sialic acid. The dominant saccharide in this milk sample was lactose, but it also contained isoglobotriose (Glc(alpha1-3)Gal(beta1-4)Glc) as well as a variety of sialyl oligosaccharides. The sialyl oligosaccharides were separated from neutral saccharides by anion exchange chromatography on DEAE-Sephadex A-50 and successive gel chromatography on Bio Gel P-2. They were purified by high performance liquid chromatography (HPLC) using an Amide-80 column and characterized by 1H-NMR spectroscopy. Their structures were determined to be those of 3'-sialyllactose, 6'-sialyllactose, monofucosyl monosialyl lactose (Neu5Ac(alpha2-3)Gal(beta1-4)[Fuc(alpha1-3)]Glc), sialyl lacto-N-neotetraose c (LST c), galactosyl monosialyl lacto-N-neohexaose, galactosyl monofucosyl monosialyl lacto-N-neohexaose and three novel oligosaccharides as follows: Neu5Ac(alpha2-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)Glc, Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc, and Neu5Ac(alpha2-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-3)Gal(beta1-4)Glc. The higher oligosaccharides contained only the type II chain (Gal(beta1-4)GlcNAc); this finding differed from previously published data on Asian elephant milk oligosaccharides. 相似文献
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Histochemistry of the development of the digestive system of Siberian sturgeon during early ontogeny 总被引:1,自引:0,他引:1
E. Gisbert § M. C. Sarasquete † P. Williot ‡ F. Castelló-Orvay 《Journal of fish biology》1999,55(3):596-616
At hatching, the yolk-sac matrix of Siberian sturgeon Acipenser baeri contained neutral glycoconjugates, glycogen, proteins rich in arginine, lysine, tyrosine, cysteine and cystine, glycoproteins containing mannose (Man) and/or glucose (Glc), N -acetyl-D-galactosamine (GalNAc), L-fucose (Fuc), sialic acid and/or N -acetyl-D-glucosamine (GlcNAc) residues, as well as neutral and acidic lipids. Buccopharyngeal and anterior oesophageal goblet cellls produced a combination of neutral and acid sialoglycoproteins, while those from the posterior oesophagus secreted only neutral glycoproteins; both types of secretions contained tryptophan and -S-S- groups and were unreactive to lectin techniques. Most intestinal goblet cells secreted mainly carboxylated and sulphated sialoglycoproteins with some rests of neutral glycoconjugates, while few of them produced only acid or neutral glycoproteins. Intestinal glycoproteins were rich in GalNAc, GlcNAc and sialic acid residues. Close relationships between digestive enzymes and morphological development of digestive organs were observed. Histochemistry of enzymes revealed that just after hatching, alkaline and acid phosphatase, ATP -ase and non-specific esterase activities were detected in the yolk sac. From the onset of exogenous feeding to the juvenile stage (30 days post-hatch), an enhancement of enzymatic activities was observed, as alkaline and acid phosphatase, ATP -ase, aminopeptidase M and nonspecific esterase sharply increased. However, lipase activity decreased in the liver and brush border of enterocytes by 13–14 days post-hatch. Two types of lipase were detected in the alimentary canal, a non-pancreatic lipase that was secreted in the cardiac stomach by gastric glands, and a pancreatic lipase, which activity was mainly detected in the brush border of the intestinal epithelium. 相似文献
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Okamura M Yokoyama N Wickramathilaka NP Takabatake N Ikehara Y Igarashi I 《Experimental parasitology》2005,110(4):406-411
The present study investigated the involvement of host sialic acids in the erythrocyte infection by two equine Babesia parasites, Babesia equi and Babesia caballi. We observed that the in vitro growth of both parasites is influenced by the removal of sialic acids from the surface of equine erythrocytes (RBC). When the parasites were cultured with neuraminidase (Nm, EC 3.2.1.18)-treated RBC, in which alpha2-3-linked sialic acid residues were removed from four membrane proteins of the RBC, B. caballi showed a significant inhibition of the erythrocyte invasion, while the intracellular development of B. equi seemed to be significantly affected. The possible involvement of host sialic acid in the erythrocyte invasion by B. caballi was also supported by a significant reduction in the parasite growth accompanied by an increased number of extracellular merozoites after the addition of exogenous 3'-sialyllactose (Neu5Acalpha(2-3)Galbeta(1-4)Glc) into the culture. These results suggest that the alpha2-3-linked sialic acid residues on host RBC play important roles in the erythrocyte infections by B. caballi and B. equi. 相似文献
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Betaine prevents loss of sialic acid residues and peroxidative injury of erythrocyte membrane in ethanol-given rats 总被引:1,自引:0,他引:1
To evaluate chronic ethanol toxicity on erythrocyte membrane and preventive action of betaine as a methyl donor, 24 male Wistar albino rats were divided into three groups: control, ethanol and ethanol plus betaine group. Animals were fed 60 ml diet per day for two months. Rats in the ethanol group were fed ethanol 8 g/kg/day. The ethanol + betaine groups were fed ethanol plus betaine (0.5% w/v). After two months, all animals were killed. Malondialdehyde (MDA) and sialic acid (SA) levels were determined in plasma samples. Osmotic fragility tests were performed on whole blood samples and erythrocyte membrane thiol contents were determined using membrane suspensions. Plasma MDA levels in ethanol-given rats were increased significantly compared to the control group of rats (p < 0.05). MDA in the betaine group was significantly lower than that in the ethanol group (p < 0.05). Erythrocyte membrane thiol contents in ethanol group were decreased compared with those of the control group (p < 0.05). Thiol contents were increased slightly after betaine therapy, but this increase was not statistically significant (p > 0.05). Plasma sialic acid levels in the ethanol group were significantly higher than in the control group (p < 0.05). Sialic acid was decreased in the betaine group compared to the ethanol group (p < 0.05). In the osmotic fragility test, we observed that chronic ethanol consumption increased erythrocyte hemolysis. Betaine protected against ethanol-induced hemolysis. Our findings show that chronic ethanol administration affects erythrocyte membrane properties and this may be related to oxidative stress. Betaine protects erythrocyte membrane alterations against chronic ethanol toxicity. Therefore betaine as a nutritional agent, may protect ethanol induced clinical problems associated with membrane abnormalities. 相似文献
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The inhibitory potency of both muramic acid (MurAc) and N-acetylmuramic acid (MurNAc) on various legume lectins, including Glc/Man- and Gal/GalNAc-specific lectins, was investigated by a haemagglutination inhibition technique. Data indicated that many lectins, especially those specific for Glc/Man, specifically interact with MurAc and MurNAc often to a greater extent than with other monosaccharides and their derivatives, such as N-acetylglucosamine (GlcNAc) and sialic acid. Glc/Man-specific lectins were also shown to interact with the muramyl-dipeptide MurNAc-D-Ala-D-isoGln. These interactions could explain why various lectins readily agglutinate some bacterial strains of which cell walls contain peptidoglycans with high amounts of MurNAc. 相似文献
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A E Udoh 《Acta physiologica Hungarica》1991,78(3):265-273
The sialic acid content of erythrocytes of three different AB0 blood groups have been studied. The sialic acid contents of erythrocyte membranes containing 300 mg protein were determined and compared. Groups 0 (Rhesus negative), AB (both Rhesus negative and positive), and B (Rhesus negative) blood differed significantly (p less than 0.05) in total sialic acid content and in the distribution of sialic acid between sialoglycoproteins and other membrane components. Membrane materials containing 300 mg total protein showed sialic acid contents of 52.73 +/- 2.2 mumol sialic acid for group 0 (Rhesus negative) 34.77 +/- 1.16 mumol for group AB (Rh negative), 32.88 +/- 1.52 mumol for AB (Rh positive) and 21.23 +/- 0.84 mumol for B (Rh negative). In group 0 (Rh. neg.) membranes 39.4 +/- 1.4% of the total sialic acid was associated with the sialoglycoproteins. The percentage of sialic acids associated with sialoglycoproteins in other erythrocyte membranes were 77.7 +/- 1.3% for group B, and 55.6 +/- 1.0% and 56.4 +/- 1.8% for group AB (Rh. negative) and (Rh. positive) respectively. The changes appear to be independent of the Rhesus grouping but dependent on the AB0 grouping since membranes of the two Rhesus types of group AB had identical total sialic acid and sialoglycoproteins sialic acids. The sialic acid densities in sialoglycoproteins also differed from one erythrocyte type to another. Group 0 (Rh. negative) membrane sialoglycoproteins had sialic acid density of 140.5 +/- 3.1 nmol/mg compared to 71.7 +/- 1.2 nmol/mg for group B and 128.1 +/- 2.2 and 124.5 +/- 4.0 nmol/mg for group AB Rhesus negative and Rhesus positive respectively.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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C Sarasquete E Gisbert L Ribeiro L Vieira M T Dinis 《European journal of histochemistry : EJH》2001,45(3):267-278
Epidermal, branchial and digestive mucous cells, and the gastric glands of larvae/postlarvae (from hatching until 45 days posthatching) of three fish species (two teleostean and a chondrostean) were investigated using conventional histochemical methods (periodic acid schiff -PAS-, diastase-PAS; alcian blue pH 0.5, 1 and 2.5) in order to distinguish neutral and acidic (carboxylated and sulphated) glycoconjugates, as well as bromophenol blue reaction for identification of proteins. Additionally, the presence and distribution of sugar residues in the oligosaccharide side chains of glycoconjugates were investigated using horseradish peroxidase (HPR)-conjugated lectins (Con A, DBA, WGA and UEA-I). Most mucous cells (digestive, epidermal and branchial) of Siberian sturgeon, Acipenser baeri, sea bream, Sparus aurata and Senegal sole, Solea senegalensis larvae were PAS- and alcian blue- (pH 2.5 and 0.5) positive, with small variations between organs/tissues and species. Bromophenol blue reaction (general proteins) was positive in a minority of the mucous cells, usually in those cells which were PAS-negative. Proteins rich in sulphydryl (-SH) and/or disulphide (-S-S-) groups related with the glycoprotein nature of the glycoconjugates present in mucous cells were also observed. Epidermal, branchial and digestive mucous cells of all studied larvae did not contain glycogen or lipids. Con A lectin staining was negative in all mucous cells types of sea bream and sole, but oesophageal mucous cell of sturgeon were reactive to different lectin reactions, suggesting the presence of mannose -Man- and/or glucose -Glc-, L-fucose -Fuc- ; N-acetyl-D-galactosamine -GalNAc-, as well as N-acetyl-D-glucosamine- GlcNAc - and/or sialic acid -NANA- residues. Digestive mucous cells of all studied larvae were positive to WGA and DBA lectins. Epidermal and branchial mucous cells of sea bream and sole were Con A, DBA and UEA-I unreactive. However, mucous cells of sturgeon larvae were stained with UEA-I lectin. Gastric glands appear very early in sturgeon stomach larvae development (between 5-6 days posthatching) but rather late (around 40 days) during the ontogeny of sole and sea bream larvae. These glands contain neutral glycoproteins with Man and/or Glc, Fuc, GlcNAc- and/or sialic acid and rich in GalNAc- sugar residues, as well as proteins moderately rich in arginine, and others particularly rich in tyrosine and tryptophan. 相似文献
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Soluble suppressor factor (SSF) is a recently purified human lymphokine produced by peripheral blood lymphocytes (PBL) in serum-free medium as a likely consequence of an autologous mixed lymphocyte reaction. Immunoregulatory actions of SSF include suppression of: polyclonal B cell activation, proliferative responses of normal PBL, and natural killer (NK) and antibody-dependent cellular cytotoxicity. We examined the ability of the monosaccharides fucose (Fuc), galactose (Gal), glucose (Glc), and mannose (Man) to reverse SSF-mediated suppression of NK activity. Fuc and Gal can partially or completely reverse SSF-mediated suppression at four effector:target cell ratios. Man and Glc were unable to significantly reverse SSF-mediated suppression. Fuc or Gal was added to PBL at various times after addition of SSF. SSF-mediated suppression of NK cytotoxicity becomes irreversible with respect to these monosaccharides during the first 24 hr of PBL exposure to SSF. To explore the mechanism behind this block of SSF-mediated suppression. Fuc or Gal (50 mM) was cultured with PBL for 24 hr before addition of SSF, or with SSF for 24 hr before addition to PBL. Our experiments indicate that SSF is directly interacting with these monosaccharides, and may function by recognizing specific sugar moieties on the surface of effector cells. 相似文献
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Fagerberg D Angström J Halim A Hultberg A Rakhimova L Hammarström L Borén T Teneberg S 《Glycobiology》2009,19(2):182-191
The "Le(b) mouse" was established as a model for investigations of the molecular events following Le(b)-mediated adhesion of Helicobacter pylori to the gastric epithelium. By the expression of a human alpha-1,3/4-fucosyltransferase in the gastric pit cell lineage of FVB/N transgenic mice, a production of Le(b) glycoproteins in gastric pit and surface mucous cells was obtained in this "Le(b) mouse," as demonstrated by binding of monoclonal anti-Le(b) antibodies. To explore the effects of the human alpha-1,3/4-fucosyltransferase on glycosphingolipid structures, neutral glycosphingolipids were isolated from stomachs of transgenic alpha-1,3/4-fucosyltransferase-expressing mice. A glycosphingolipid recognized by BabA-expressing H. pylori was isolated and characterized by mass spectrometry and proton NMR as Fuc alpha 2Gal beta 3(Fuc alpha 4)GalNAc beta 4 Gal beta 4 Glc beta 1Cer, i.e., a novel Le(b)-like glycosphingolipid on a ganglio core. In addition, two other novel glycosphingolipids were isolated from the mouse stomach epithelium that were found to be nonbinding with regard to H. pylori. The first was a pentaglycosylceramide, GalNAc beta 3 Gal alpha 3(Fuc alpha 2)Gal beta 4 Glc beta 1Cer, in which the isoglobotetrasaccharide has been combined with Fuc alpha 2 to yield an isoglobotetraosylceramide with an internal blood group B determinant. The second one was an elongated fucosyl-gangliotetraosylceramide, GalNAc beta 3(Fuc alpha 2)Gal beta 3GalNAc beta 4Gal beta 4 Glc beta 1Cer. 相似文献