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1.
油麻藤种子中凝集素的纯化及性质的研究   总被引:4,自引:0,他引:4  
中药常春油麻藤种子中含有A型血专一性的凝集素(MSL)。该凝集素可经盐析、离子交换及凝胶过滤进行纯化,当其浓度为0.49μg/ml时就能凝集人A型血细胞,对人类B、O型及兔红细胞无作用。Gal,GalNAc和胃粘蛋白对MSL的凝血活性有强抑制作用。MSL含中性糖5.7%,凝胶过滤测得分子量为131800,SDS-PAGE测得分子量为66000和33000,表明MSL可能由两个不同亚基组成。MSL还  相似文献   

2.
大瓶螺蛋白腺经磷酸盐缓冲液抽提、硫酸铵分级沉淀、SephadexG-100和Sepharose4B凝胶过滤,可获得在不连续PAGE(pH4.3和pH8.9)上显示单一蛋白质染色带的大瓶螺凝集素(AGL).该凝集素对人血红细胞无血型专一性,但对A型血红细胞的凝集作用最强.AGL的血凝活力可被乳糖或半乳糖所抑制.AGL分子中的中性糖含量为0.24mg/mg蛋白质.用SDS-PAGE法测得其亚基分子量为15000,且只有一种亚基.AGL中Cys和Phe的含量较高,并较耐热.  相似文献   

3.
大瓶螺凝集素的分离纯化及部分性质研究   总被引:2,自引:0,他引:2  
大瓶螺蛋白腺经磷酸盐缓冲液抽提、硫酸铵分级沉淀、Sephadex G-100和Sepharose 4B凝胶过滤,可获得在不连续PAGE(pH4.3和pH8.9)上显示单一蛋白质染色带的大瓶螺凝集素(AGL)。该凝集素对人血红细胞无血型专一性,但对A型血红细胞胞的凝集作用最强。AGL的血凝活力可被乳糖或半乳糖所抑制。AGL分子中的中性糖含量为0.24mg/mg蛋白质。用SDS-PAGE法测得其亚基分  相似文献   

4.
棉花凝集素的纯化及性质研究   总被引:7,自引:0,他引:7  
抗枯萎、抗黄萎病的棉种的浸取液,经硫酸铵分级,纤维素柱、亲和层析后,再经分子筛过滤,获得在PAGE、SDS-PAGE或HPLC柱上均呈现单一蛋白带的棉花凝集素。该凝集素只凝集兔红细胞,对人A、B或O型血细胞均不凝集,其凝集活性可被半乳糖或猪甲状腺球蛋白等所抑制。棉花凝集素在65℃加热5 m in,即丧失全部凝集活性;它的凝集活性强烈地依赖于Ca2+ ;Mn2+ 对活性也有促进作用,Mg2+ 则无作用。经凝胶过滤或SDS-PAGE测定,凝集素的分子量为63000,N-末端氨基酸为Val。凝集素含有1.5% 的中性糖,是一种促有丝分裂原,细胞转化率为50.3% 。  相似文献   

5.
白茯苓凝集素的研究   总被引:1,自引:0,他引:1  
以中药白茶苓为材料,采用盐析、离子交换及凝胶过滤可纯化出快速蛋白液相谱(FPLC)纯的白茯苓凝集素(SLL)。当其浓度为0.25μmg/mL时就能凝集兔红细胞,对人类A,B,O型血细胞无作用。甲状腺球蛋白、卵粘蛋白和胃蛋白对SLL的凝血活性有强抑制作用,而常见糖类对其无明显抑制作用。SLL含中性糖12.5%,凝胶过滤及FPLC测得分子量为31000,亚基分子量为8000,表明SLL可能由四个相同亚  相似文献   

6.
厚果鸡血藤凝集素的纯化及性质   总被引:2,自引:0,他引:2  
从厚果鸡血藤(MiletiapachycarpaBenth.)的种子中分离纯化出一种具强凝集活性和强促有丝分裂原的凝集素。种子经磨粉、浸取、硫酸铵分级、DEAESepharose离子交换和SephadexG100分子筛层析,即可获得在PAGE和SDSPAGE上均呈现单一蛋白染色带的凝集素纯品,分子筛层析测得分子量为40700,SDSPAGE测得亚基分子量为19800;含有178%的中性糖。氨基酸组成分析表明,该凝集素富含Asp、Glu、Thr、Ser和Leu,同时含有4个Trp,当凝集素浓度为0.48μg/mL时,即可凝集兔红细胞;对人A、B和O型血细胞都能发生凝集,故无血型专一性;其凝集兔红细胞的凝集活性,不能被常见糖类抑制,但可被甲状腺球蛋白、胃粘蛋白和卵粘蛋白所抑制;其凝集活性强烈地依赖于Ca2+的存在,但Mg2+、Mn2+、Zn2+对其凝集活性全无促进作用;该凝集素是一种强促有丝分裂原,对人外围血中淋巴细胞的转化率高达843%,细胞分裂比率可达78%。  相似文献   

7.
黄精凝集素Ⅱ的纯化及部分性质研究   总被引:5,自引:0,他引:5  
囊丝黄精(PolygonatumcyrtonemaHua.)的根状茎,经浸取、用硫酸铵分级沉淀、猪甲状腺球蛋白-Sepharose4B柱亲和层析、CM-Sepharose柱离子交换层析和SephadexG-100凝胶过滤,可以分离纯化出黄精凝集素Ⅱ(PCLⅡ).纯化的PCLⅡ在聚丙烯酰胺凝胶电泳中显示单一蛋白染色带;在快速高效液相色谱中亦为单一蛋白峰,经分子筛层析测得分子量为15.9kD,最大紫外吸收值在278nm,PCLⅡ只凝集兔红细胞,当浓度为0.25μg/ml时,即可发生凝集反应,此凝集兔红细胞的能力可被D-甘露糖和猪甲状腺球蛋白所抑制.氨基酸组成分析表明PCLⅡ分子中富含酸性氨基酸,N末端为丙氨酸.经测定PCLⅡ分子中含有3个色氨酸和2.4%的中性糖.原子发射光谱分析表明,该凝集素分子中含有Mg和Ca两种金属元素.  相似文献   

8.
以中药白茯苓为材料,采用盐析、离子交换及凝胶过滤可纯化出快速蛋白液相谱(FPLC)纯的白茯苓凝集素(SLL)。当其浓度为0.25μg/mL时就能凝集兔红细胞,对人类A,B,O型血细胞无作用。甲状腺球蛋白、卵粘蛋白和胃粘蛋白对SLL的凝血活性有强抑制作用,而常见糖类对其无明显抑制作用。SLL含中性糖12.5%,凝胶过滤及FPLC测得分子量为31000,亚基分子量为8000,表明SLL可能由四个相同亚基组成。SLL对酸、碱、热均有较高的耐受性,并且是一个强促有丝分裂原。  相似文献   

9.
为了探讨凝集素在豆科植物与根瘤菌的识别过程中的作用,用DEAE-32离子交换层析和SephadexG-150凝胶过滤分离、纯化格拉姆柱花草种子凝集素(SGL),其分子量约为45kD,由两个相同的亚基组成,等电点约pH5.8,它是一种糖蛋白,含糖量约为2.6%.SGL的热稳定性强.SGL的血凝活性能被甘露糖所抑制.SGL对红细胞的凝集作用可能具有种属专一性;SGL具有强的促有丝分裂作用;荧光标记实验显示:9株能与格拉姆柱花草植株结瘤的菌株有7株能与SGL结合,6株不能与之结瘤的菌株,只有1株能与SGL结合,这表明不同根瘤菌菌株对SGL的结合能力,和它们在格拉姆柱花草上结瘤能力之间可能具有一定的生物学相关性.  相似文献   

10.
野花生豆凝集素的化学修饰与荧光光谱研究   总被引:4,自引:0,他引:4  
野花生豆凝集素(CML)经Sephadex G-200测得分子量为103.O kD。用对二甲基氨基苯甲醛(DAB)为显色剂,测得每个CML分子含有5.9个色氨酸残基。在pH5.1,含8mol/L脲的醋酸缓冲中,N-溴代丁二酰亚胺(NBS)可修饰CML分子中的5.6个色氨酸(Trp)残基,同时使CML的凝血活性完全丧失。用焦碳酸二乙酯(DEPC)和N-乙酰顺丁烯酰胺(NEM)分别修饰CML的组氨酸残  相似文献   

11.
番茄成熟果实经抽提上鸡卵类粘蛋白(OM)-Sepharose 4B亲和层析柱分离纯化制得番茄凝集素。制品在SDS—聚丙烯酰胺凝胶电泳上呈一条带,表观分子量为205,000。Sephadex G200凝胶过滤行为呈单一吸收峰,分子量为180,000。等电聚焦测得等电点为7.6和9.4。它能使人和多种动物红细胞,以及某些培养细胞凝集,但效价不同。N—乙酰葡萄糖胺寡聚糖是其专一性结合糖。  相似文献   

12.
黑色菜豆(phaseolussp.)种子中含有对人A型血专一凝集的凝集素。用猪胃粘蛋白-Sepharose 4B作亲和吸附剂和Sephadex G-200凝胶过滤,可以纯化这种凝集素。纯化的凝集素在pH8.9,Tris-EDTANa_2-borate缓冲液的PAGE中,呈现单一蛋白带;酚-硫酸法测得总糖含量为3.22%。在SDS-PAGE中发现其分子由两种亚基所组成,亚基分子量分别为38,000和35,000。当凝集素浓度分别为0.98μg/ml和1.95μg/ml时能强烈地凝集人A型和AB型血细胞。在凝集素浓度高达500μg/ml时,B型血细胞能发生弱凝集反应,但对O型血和兔红细胞则完全不发生凝集反应。其凝集活性可被GalNAC、L-Fuc、猪甲状腺球蛋白和卵粘蛋白所抑制。该凝集素对人外周血中淋巴细胞的转化率达80%,细胞分裂比率高达37.1%;氨基组成分析表明,凝集素分子中Asp和Glu含量较高,而cys和Met含量很低。  相似文献   

13.
用猪甲状腺球蛋白-Sepharose 4B作亲和吸附剂,再经Sephadex G-100凝胶过滤,可以从岩豆种子中纯化出岩豆凝集素(MDL)。该凝集素可以凝集人类A、B、O型血细胞和兔红细胞,纯化的MDL凝集兔红细胞的能力可被D-松三糖、邻硝基-苯酚-D-半乳糖和N-乙酰半乳糖胺抑制,甘露糖也有弱的抑制作用。纯化的MDL在PAGE和SDS-PAGE上均显现单一蛋白质染色带,经Schiff’s试剂染色证明为糖蛋白;以酚-硫酸法测得其中性糖含量为6.0%;SDS-PAGE测得亚基分子量为32 000;Sephadex G-100分子筛柱测得其分子量为63 800;等电聚焦电泳显示其等电点为5.1;氨基酸组成分析表明其中Asp、Glu、Phe含量较高,但不含有Pro、Tyr。MDL也是一个强促有丝分裂原,对人外周血淋巴细胞转化率可达81.2%,细胞分裂比率达14.8%。  相似文献   

14.
A lectin (LEL) was isolated from the fresh fruiting bodies of the shiitake mushroom Lentinula edodes by a combination of gel filtration chromatography on Sephadex G-150 and affinity chromatography on an N-acetyl-Dgalactosamine-Sepharose 4B column. Its molecular mass, as determined by gel filtration, was estimated to be 71, 000 Daltons and its structure is homotetrameric with subunit molecular weight of approximately 18,000 Daltons. LEL agglutinated non-specifically red blood cells from the human ABO system as well as rabbit erythrocytes and in haemagglutination inhibition assays, exhibited sugar-binding specificity toward N-acetyl-D-galactosamine. EDTA had no inhibitory effect on its haemagglutinating activity, which was stable up to 70°C and was not affected by changes in pH. The lectin had no covalently-linked carbohydrate and amino acid composition analysis revealed that it contained 124 amino acid residues and was rich in tyrosine, proline, phenylalanine, arginine, glutamic acid and cysteine. LEL did not cause mortality neither was it observed to alter the morphology of key organs when administered intraperitoneally at concentrations up to 10,000 mg kg-1 body weight of mice.  相似文献   

15.
 用猪甲状腺球蛋白-对氨基苯砜乙基-交联琼脂作亲和吸附剂,可对雪山豆(Phaseolus sp.)凝集素进行亲和层析纯化。纯化后的凝集素在聚丙烯酰胺凝胶电泳中显示单一蛋白质带。Sephadex G-100凝胶层析法测得分子量为65,000道尔顿,SDS-聚丙烯酰胺凝胶电泳表明该凝集素分子仅有一个分子量为65,000道尔顿的亚基,酚-硫酸法测得总糖含量为4.6%,氨基酸组成分析表明雪山豆凝集素富含门冬氨酸,而甲硫氨酸含量甚少。该凝集素是强促有丝分裂原,对人外周血中淋巴细胞的转化率大于90%,细胞分裂比率达12%。雪山豆凝集素不仅能凝集多种动物红细胞,还能凝集人精细胞。经体外实验表明,雪山豆凝集素对人肝癌细胞有抑制作用。  相似文献   

16.
Extracts of the marine polychaetous annelid, Amphitrite ornata, agglutinate rat, rabbit, chicken and human erythrocytes and in other work have been shown to inhibit the growth of Ehrlich ascites tumors in mice. Fractionation of extracts on Sephadex G-100 gave three active fractions with molecular weights of 30 000, 54 000 and 100 000. The 30 000 dalton fraction (B) was purified 72-fold by ammonium sulfate precipitation, gel filtration and preparative disc gel electrophoresis. The purified hemagglutinin, amphitritin, was homogenous on analytical disc gel electrophoresis at four different pH values and gave a sharp boundary in sedimentation velocity ultracentrifugation. The three fractions showed paralled specificity toward rat and chicken erythrocytes, the former giving the higher titer. The purified agglutinin was active toward human blood groups A, B and O and exhibited 4-fold higher activity toward group A. The hemagglutinin titer against rat red blood cells was lowered only by N-acetylgalactosamine, the terminal sugar residue of the group A determinant. None of the saccharides tested inhibited agglutination of chicken erythrocytes. Hemagglutinin activity was insensitive to dialysis or treatment with EDTA. The activity was not affected by digestion with trypsin or pronase, but was destroyed by phenol extraction. Analytical disc gel electrophoresis showed one protein band with high anodal mobility at pH 8.5, which was not affected by proteolytic enzymes but was removed by phenol. Activity was unaffected by heating at 70 degrees C for 30 min but was destroyed by similar treatemtn at 85 degrees C. Activity was at a maximum at pH 7-9 and decreased reversibly down to pH 4 at which point it was irreversibly inactivated. The higher molecular weight agglutinin (A1) could be dissociated to give amphitritin by treatment with 6M urea of precipitation in 55% (NH4)2SO4. This dissociation was not reversed by dialysis. Amphitritin is a glycoprotein with a molecular weight determined by gel filtration of 30 000 and by approach to equilibrium sedimentation of 32 000. Amino acid analysis showed a preponderance of aspartic and glutamic acids and relatively large amounts of glycine, proline, alanine, valine and cysteine. The carbohydrate moeity which represented 12.8% of the molecule, contained mannose, galactose, glucosamine and sialic acid. Amphitritin is the first hemagglutinin to be isolated from a polychaetous annelid.  相似文献   

17.
用猪胃粘蛋白-Sepharose 4B作亲和吸附剂,可从野花生豆(Crotalarta mucronata)的种子中分离纯化出对人类A型血专一凝集的凝集素。该凝集素可用pH30.,Gly-HCl-1mol/L NaCl溶液解吸附。纯化的凝集素在PAGE或SDS-PAGE中均显示单一蛋白带,表明凝集素分子内只有一种亚基。用SDS-PAGE测得其亚基分子量为49,000。氨基酸组成分析表明,该凝集素富含甘氨酸和谷氨酸,不合甲硫氮酸。纯化的野花生豆凝集素(简称CML)含有4.11%的中性糖。它对人A型血细胞有强烈凝集作用,对AB型血有弱凝集作用,但对B型和O型血均不凝集。其对A型血细胞的凝集作用可被N-乙酰半乳糖胺抑制,但对AB型血则无抑制作用。CML是一个促有絲分裂原,对人外周血中淋巴细胞有促有絲分裂作用。  相似文献   

18.
A lectin with N blood group specificity was isolated from Vicia graminea seeds. This lectin was purified from a crude extract by precipitation with ammonium sulfate, DEAE-cellulose chromatography and Sephadex G-150 gel filtration. Purification steps were followed by increase of specific activity. Its homogeneity was demonstrated by polyacrylamide gel electrophoresis, immunoelectrophoresis, electrofocusing and ultracentrifugation. This lectin is an acid glycoprotein with 7.3% carbohydrate, a high percentage of serine and contains no sialic acid. The native lectin has a molecular weight about 100 000 and dissociates into four subunits of 25 000 as shown by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Preliminary hemagglutination inhibition has shown that the lectin was not inhibited by any of the monosaccharides contained in N blood group substances; however it was inhibited by the erythrocyte membrane major glycoprotein and the tryptic fragments obtained from erythrocytes.  相似文献   

19.
芦荟凝集素的分离、纯化和部分性质的研究   总被引:5,自引:0,他引:5  
新鲜芦荟叶(Aloe vera L.var.chinensis(Haw.)Berger)于室温用低浓度NaCl溶液提取。离心和透析后,经N-乙酰氨基葡萄糖0Sepharose 4B亲和层析,分离纯化出芦荟凝集素(ACL)。用SephadexG-100测表观分子量为35KD,SDS-PAGE出现两条色带;染色 ;宽带和较浅的罕带。亚基分子量分别为15KD和20KD。能专一性凝集兔血细胞和人血红细胞  相似文献   

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