Purification and Partial Characterization of a Lectin from the Bark of Japanese Elderberry (Sambucus sieboldiana)

A lectin [Sambucus sieboldiana agglutinin (SSA)] was purifiedfrom the twigs of Sambucus sieboldiana by repeated affinitychromatography on fetuin-Sepharose. SSA had a molecular weight(Mr) of approximately 160 K on gel filtration and consistedof two types of subunit of which the molecular weights rangedfrom 31 to 37 K. SSA agglutinated human erythrocytes irrespectiveof their blood type and the hemagglutination was inhibited bya very low concentration of Neu5Ac(2-6)lactose, suggesting thatSSA has a carbohydrate-binding specificity similar to that ofthe lectin previously isolated from the bark of S. nigra (SNA).However, the Ouchterlony double-diffusion analysis of theselectins with an antibody raised against SSA showed that SSAwas immunologjcally related but not identical to SNA. (Received January 17, 1989; Accepted June 19, 1989)     

Purification and Partial Characterization of a Lectin from Phaseolus vulgaris

A method is presented for the isolation of a lectin from a Brazilian cultivar of the common bean (Phaseolus vulgaris L.), through extraction in acidic (pH 4.2) medium, fractionation with ammonium sulfate, and chromatography on DEAE-cellulose. The lectin was shown to be homogeneous by gel electrophoresis and isoelectric focusing.     

三齿草藤(Vicia bungei Ohwi)凝集素的亲和层析纯化及其部分性质的研究

用Sephadex G-100或猪甲状腺球蛋白-对氨基苯砜乙基-交联琼脂作亲和吸附剂,均可从三齿草藤(Vicia Bungei Ohwi)的种子中分离纯化出三齿草藤凝集素。该凝集素经连续或不连续系统聚丙烯酰胺凝胶电泳均显示出单一蛋白带;糖蛋白染色法证实为糖蛋白;SDS-聚丙烯酰胺凝胶电泳测定其分子量为24,600,凝集素浓度为1.95微克/毫升时就能凝集兔红细胞;但对人ABO型血细胞不发生凝集作用;其对兔红细胞的凝集作用可被D-Man、D-GlcNA和D-GIC所抑制;它也是一种促有丝分裂原。     

香灰菌凝集素的纯化及其部分性质

香灰菌菌丝体经磷酸缓冲液抽提、20%-70%饱和浓度的硫酸铵沉淀、DEAE-Cellulose和SephadexG-100柱层析纯化得到香灰菌凝集素(Hypoxylonsp.lectin,简称HSL)。HSL经PAGE检测为单一蛋白条带,SDS-PAGE测得其亚基分子量为15.9kD。过碘酸-Schiff染色法表明HSL为一种糖蛋白,糖基的含量为15.5%,β-消去反应测得其糖和蛋白质的连接键为O-型糖肽键。HSL能凝集多种动物红细胞和人的红细胞,在所测试的红细胞中,对兔红细胞的凝集作用最强。HSL对热较敏感,经50°C处理10min,其凝集活性明显降低,其在碱性环境中较稳定,而在酸性环境中较不稳定。HSL的凝集活性受Al3+、Fe3+、Ca2+和Zn2+等阳离子的影响。对鼠红细胞的凝集作用可被半乳糖和乳糖所抑制。     

Purification and Characterization of Cotton Lectin

Defatted seeds of wilt-disease resistant were extracted overnight with PBS at 4 ℃. After centrifugation. 90% saturated (NH4)2SO4 was added to the supernatant. The precipitates were dialysed against H2O, then lyophilized. The purified lectin was obtained by DEAE-cellulose ion-exchange chromatography, Sephadex G-100 filtration, and Sepharose 4B-Hog thyroglohulin affinity chromatography. The activity of the lectin was tested with fresh rabbit erythrocyte in each step of the procedure, and the active part was collected. This sample demonstrated single band on PAGE, SDS-PAGE and HPLC. The lectin was a glycoprotein. It contained 1.5% of neutral saccharide and its molecular weight was 63000 determined by Sephadex G-100 filtration. The N-terminal amino acid of the lectin was Val. The lectin showed no specific agglutination with any type of human erythrocytes. The hemagglutinaition activity could be inhibited by galactose and hog thyroglobilin, and depended on Ca2+, Mn2+, especially on Ca2+, not Mg2+. Its biological activity lost at 65 ℃ for S min. This lectin is used as a mitogen for human peripheral blood lymphocytes.     

Purification and characterization of Robinia pseudoacacia seed lectins. A re-investigation.

Two lectins, RPA 1 and RPA 3, were purified from Robinia pseudoacacia seeds. These two lectins differ in their physicochemical and biological properties. By analytical ultracentrifugation the Mr values of RPA 1 and RPA 3 were estimated to be 59,000 and 105,000 respectively. From SDS/polyacrylamide-gel-electrophoresis data it was estimated that RPA 1 consisted of two subunits of Mr 34,000, and RPA 3 of two types of subunits (Mr 30,500 and 29,000). RPA 1 and RPA 3 were found to be glycoproteins of comparable amino acid composition. RPA 1 was the more highly glycosylated molecule (11.6% versus 4.3%). The carbohydrate-specificity of RPA 1 appears to be complex. RPA 3 was inhibited by N-acetyl-D-galactosamine and human alpha-glycoproteins. Both lectins exerted a mitogenic effect on human peripheral-blood lymphocytes. Concentrations between 0.5 and 1 microgram of RPA 3/ml gave optimal proliferative responses, whereas for RPA 1 concentrations higher than 10 micrograms/ml were needed for these responses.     

Purification and Characterization of Sophora flavescens Lectin

A lectin with strong hemagglutination activity was isolated from roots of Sophora flavescens Ait. by extraction, fractionation with (NH 4) 2SO 4, ion-exchange chromatography on DEAE-Sepharose and followed by gel filtration on Sephadex G-150 and HPLC assay. The purified lectin showed a single protein band on PAGE and SDS-PAGE . The molecular weight of S. flavescens lectin was 32 kD when SDS-PAGE and Sephadex G-100 was used. The lectin agglutinated rabbit red blood cells at 0.97 μg/mL and showed no specific agglutination with any type of human erythrocytes. The hemagglutination activity could be inhibited by mannose and levulose and slightly by glucose and maltose. The SFL contained 2.89% neutral saccharide. It could inhibit apparently the growth of the mycelium of Gibberlla saubinetii （Mont.） Sacc.,Piricularia oryzae Cav. and Fusarium vasinfectum Atk. at the dosage of 62 μg. It was determined by Edman that the sequence of the N-terminal thirty amino acids was: T/A/VDXLXFTFSDFDPNGEDLLFQGDAHVTSNN.     

狗脊蕨凝集素的分离纯化与部分性质

狗脊蕨叶组织经磷酸缓冲液抽提,硫酸铵分级沉淀,DEAE-Sepharose(fast flow)离子交换层析和Sephadex G-100分子筛层析,获得具有凝集活性的狗脊蕨凝集素.用PAGE和SDS-PAGE检测均显示1条蛋白带,提示狗脊蕨凝集素分子只有一个亚基.Sephadex G-100层析和SDS-PAGE测得其相对分子质量在20 kD左右.该凝集素对红细胞有种属专一性,其凝集活性可被木糖、卵粘蛋白所抑制.狗脊蕨凝集素对热相当稳定,以100℃温度加热10 min仍保存部分凝集活性.凝集活性不依赖于金属离子Mg2 、Ca2 、Mn2 .它的中性糖含量为3.1%,Phe含量较高,His含量较低,不含Tyr和Pro.     

苦参凝集素的分离纯化及部分性质研究

从苦参 (SophoraflavescensAit.)根浸出液经硫酸铵分级 ,得苦参凝集素 (SFL)粗品 ,再经DEAE_Sepharose、SephadexG_1 5 0和HPLC层析 ,获得具有强凝集活性的SFL样品 ,用PAGE和SDS_PAGE检测均为单一蛋白染色带。SDS_PAGE显示SFL分子仅有一条肽链 ,SephadexG_1 0 0和SDS_PAGE测得其分子量均为 32kD。当SFL浓度为 0 .97μg/mL时能凝集兔红细胞 ,无血型专一性 ,其凝血活性可被甘露糖和果糖抑制 ,麦芽糖和葡萄糖有弱的抑制作用 ,凝集素分子含有 2 .89%的中性糖 ;当SFL量为 6 2 μg时 ,对棉花枯萎病菌 (FusariumvasinfectumAtk .)、小麦赤霉病菌(Gibberllasaubinetii (Mont.)Sacc .)和水稻稻瘟病菌 (PiriculariaoryzaeCav)菌丝体的生长发育有明显地抑制作用。用Edman法在蛋白测序仪上测出SFL的N端肽链 30个氨基酸的排列顺序为 :T/A/VDXLXFTFSDFDP NGEDLLFQGDAHVTSNN。     

刺槐叶绿素荧光特性的研究

用L I-6400光合仪对黄土高原常用造林树木刺槐的叶绿素荧光动力学曲线、淬灭分析、荧光光曲线以及荧光AC I曲线等生理特性进行测定和对比分析.结果表明:随着光照时间的加大,刺槐的荧光参数ETR、qP和N PQ逐渐上升并在21 m in左右达到稳定,与光合同步.最大荧光产量F m在叶片转入黑暗后逐渐上升,在44 m in左右趋于稳定;N PQ则开始下降,在35 m in左右达到稳定.当光强度>200μm o l.m-2.s-1时,Ph iPS2和Ph iCO2呈线性正相关,而当光强度<200μm o l.m-2.-s 1时,Ph iPS2和Ph iCO2呈线性负相关.CO2浓度和刺槐的荧光光合速率、Ph iPS2、Ph iCO2值存在着一定的正相关关系.     

短裙竹荪(Dityophora duplicata)凝集素纯化与生化性质

短裙竹荪子实体经生理盐水抽提、硫酸铵沉淀、DEAE Sepharose和SephadexG 10 0柱层析纯化得到短裙竹荪凝集素 (Dityophoraduplicata(Bosc)Fischerlectin) ,简称DDFL .DDFL经PAGE显示单一条带 ,SDS PAGE测得其亚基分子量为 2 2 3kD ,SephadexG 10 0凝胶过滤测得分子量为 4 5 3kD ,DDFL不含中性糖 ,IEF测得其等电点为 3 92 .该凝集素对供试的 4种血型人血和兔、小牛、鸭、鸡、鲫鱼以及青蛙血红细胞具有凝集作用 ,但不凝集鳖红细胞 .它还可以凝集小鼠脾脏淋巴细胞和小鼠S180 肉瘤细胞 ,对兔红细胞的凝集作用可被乳糖、棉子糖、半乳糖、α 甲基半乳糖、β 甲基半乳糖和N 乙酰半乳糖胺所抑制 .氨基酸组成分析表明 ,DDFL含有 17种氨基酸 ,其中天冬氨酸、丝氨酸、苯丙氨酸和丙氨酸含量较高 .经测定 ,其N末端为甘氨酸 .DDFL对热、酸和碱具有一定的稳定性 ,经 6 0℃处理 10min ,可保持较高的活性 ,在pH 4 0～ 9 0范围内较稳定 ,其凝血活性依赖于Mg2 + 和Ca2 + 二价阳离子 ,Mn2 + 和Zn2 + 则无影响 .DDFL对小鼠腹腔注射的半致死量为 70 6 3mg kg .     

Purification and Characterization of the Millettia pachycarpa Lectin

A lectin with strong hemagglutinating and mitogenic activity was isolated from the seeds of Millettia pachycarpa Benth. by extraction, fraction with (NH4)2 804, ion-exchange chromatography on DEAE-Sepharose and followed by gel filtration on Sephadex G-100. The purified leetin showed a single protein band on PAGE and SDS-PAGE, and exhibited a molecular weight of 40 700 by gel filtration and subunit of 19 800 on SDS-PAGE. It contained 1.78% neutral saceharide and enriched Asp, Glu, Thr, Set, Leu and also contained 4 Trp per molecule. It agglutinated rabbit red cell at 0.48 μg/mL and A,B,O types of blood. The reaction could be inhibited by thyroglobulin, muein gastrie and ovomucin, but not by saceharide. The hemagglutination was strongly dependant on Ca2 +, but was not enhanced by Mg2 + ,Mn2 + ,Zn2 +; The lectin was a strong mitogen for human peripheral blood lymphoeytes, showing a transformation rate and mitotic index of 84.3 % and 7.8 %,respectively.     

Purification and Characterization of Lectin from Lathyrus sativus

Lectin has been isolated and purified from Lathyrus sativus using ammonium sulphate precipitation followed by affinity chromatography. The molecular weight as determined by HPLC was found to be 42kD. The lectin is a tetramer, consisting of two types of subunits of which the heavier subunit consists of 2 polypeptides of mol wt of about 21 kD and 16 kD while the smaller subunits consists of two polypeptides of about 5kD as revealed by SDS-PAGE. The most potent sugar inhibitor of the Lathyrus lectin was found to be α-methyl D-mannoside. The N-terminal amino acid sequence was similar to that of pea lectin sequence.     

格拉姆柱花草(Stylosanthes gracilis H.B.K.cv.Graham)凝集素的纯化与性质研究

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．     

旋扭山绿豆（Desmodiumintortum）凝集素的纯化与特性

旋扭山绿豆（Ｄｅｓｍｏｄｉｕｍｉｎｔｏｒｔｕｍ）凝集素的纯化与特性彭建宗,程双奇,陈兆平,莫熙穆（华南师范大学生物系,广州５１０６３１）关键词凝集素;旋扭山绿豆;亲和层析;糖蛋白豆科植物和根瘤菌之间的共生结瘤固氮关系早有报道。例如大豆只被Ｂ．仰。６。...     

猪精子凝集素的纯化,性质及其作用

用胎球蛋白－Ｓｅｐｈａｒｏｓｅ亲和层析和凝胶过滤层析从精子和精浆中分离纯化了猪精子凝集素（简称ＢＳＬ）。ＢＳＬ的血凝活性只被若干糖蛋白和聚糖所抑制。ＢＳＬ的分子量为５６ｋｄ,由分子量分别为１３．６ｋｄ（β）和１６．０ｋｄ（α）的两个不同的亚基以α１β３所组成。ＢＳＬ为糖蛋白,含中性糖３．２％,不含唾液酸。用ＥＬＩＳＡ法测定猪精子中ＢＳＬ的含量及分布,表明７０％嵌入在精子膜中,２５％结合在精子表面,     

Agrobacterium tumefaciens-mediated transformation of Robinia pseudoacacia

Robinia pseudoacacia (black locust) plants were regenerated after co-cultivation of stem and leaf segments with Agrobacterium tumefaciens strain GV3101 (pMP90) that harbored a binary vector that included genes for β-glucuronidase (GUS) and hygromycin phosphotransferase. Successful transformation was confirmed by the ability of stem and leaf segments to produce calli in the presence of hygromycin, by histochemical and fluorometric assays of GUS activity in plant tissues, and by Southern blotting analysis. In this transformation system, about 2 months were required for regeneration of transgenic plants from stem and leaf segments. The frequency of transformation from stem segments was approximately 24%, and the morphology of regenerated plants resembled that of the original parental strain. Received: 2 September 1999 / Revision received: 30 November 1999 / Accepted: 4 December 1999     

Purification and Characterization of a Lectin from Crotalaria paulina Seeds

A lectin was purified from Crotalaria paulina seeds by ion-exchange and FPLC molecular exclusion chromatography. CrpL had an apparent molecular mass of 30 kDa, as determined by SDS-PAGE under non-reducing and reducing conditions. CrpL effectively agglutinated human and cow erythrocytes, and this activity was not affected by 20 mM EDTA, showing no dependence of divalent cations. Hemagglutination was inhibited by N-acetyl- D-galactosamine, D-galactose and was also inhibited by glycoproteins, fetuin and asialofetuin. The N-terminal amino acid sequence of CrpL was identical to those of other lectins from the genus Crotalaria, and amino acid composition showed high amounts of Asx and Glx, and was rich in Gly, Ala and Ser, as also reported for lectins from other Crotalaria species. CrpL inhibited the growth of Xanthomonas axonopodis pv. phaseoli and Xanthomonas axonopodis pv. passiflorae, suggesting a role of this lectin in the defense of seeds against bacterial infections.     

岩豆(Millettia dielsiana Harms ex Diels)凝集素的分离纯化和性质研究

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％。     

Purification and Characterization of Lectin from Seeds of Delonix regia

A lectin from the crude extract of seeds of Delonix regia (DRL) has been purified by ammonium sulphate fractionation followed by specific adsorption on Sephadex G-50 column and subsequent displacement with 100 mM D-glucose. The purified lectin (yield 1.41 mg g^?1 dry seed) is a hetero-tetramer of 156 kD in size, consisting of four polypeptides (M_r of 32, 36, 42 and 46 kD) as detected on SDS-PAGE. It is a thermostable protein and remains active between pH 2.0–11.0. The lectin agglutinated erythrocytes of human and other primates. The hemagglutinating activity was not affected by cations and chelating agents. Of the 23 different sugars tested for specificity, maximum inhibition of the hemagglutination was shown by D-glucose. The immunological crossreactions of DRL with monospecific antibodies against SBA, Con A, PNA, DBA and PHA-E indicate that DRL is very closely related to Concanavalin A.