共查询到20条相似文献,搜索用时 15 毫秒
1.
E. P. Vetchinkina V. E. Nikitina O. M. Tsivileva L. V. Garibova 《Applied Biochemistry and Microbiology》2008,44(1):66-72
We studied changes of the hemagglutinating activity of intracellular lectins of the basidiomycete Lentinus edodes (shiitake) at various stages of its morphogenetic development depending on erythrocyte type, growth medium, and lectin purification degree. Under certain experimental conditions, the specific lectin activity at the brown mycelium film stage exceeded the corresponding value for nonpigmented mycelium. The sensitivity of the lectins towards trypsin-treated rabbit erythrocytes was no less than a hundredfold higher than towards any other erythrocyte type studied. The general regularities of specific activity change did not depend on nutrient medium composition. With purification of intracellular shiitake lectins, their sensitivity to human erythrocytes decreased seventyfold or more, whereas their sensitivity to rabbit erythrocytes increased by the same factor. 相似文献
2.
Cammarata M Benenati G Odom EW Salerno G Vizzini A Vasta GR Parrinello N 《Biochimica et biophysica acta》2007,1770(1):150-155
A novel fucose-binding lectin, designated SauFBP32, was purified by affinity chromatography on fucose-agarose, from the serum of the gilt head bream Sparus aurata. Electrophoretic mobility of the subunit revealed apparent molecular weights of 35 and 30 kDa under reducing and non-reducing conditions, respectively. Size exclusion analysis suggests that the native lectin is a monomer under the selected experimental conditions. Agglutinating activity towards rabbit erythrocytes was not significantly modified by addition of calcium or EDTA; activity was optimal at 37 degrees C, retained partial activity by treatment at 70 degrees C, and was fully inactivated at 90 degrees C. On western blot analysis, SauFBP showed intense cross-reactivity with antibodies specific for a sea bass (Dicentrarchus labrax) fucose-binding lectin. In addition, the similarity of the N-terminal sequence and a partial coding domain to teleost F-type lectins suggests that SauFBP32 is a member of this emerging family of lectins. 相似文献
3.
研究三叶半夏内生真菌及其凝集素,旨在为半夏内生真菌及其凝集素的开发利用提供依据。对三叶半夏块茎内生真菌分离、纯化,液体发酵培养代谢产物,无水乙醇提取总蛋白,兔血红细胞检测其凝集活性,筛选出菌株gs1,其总蛋白对兔血红细胞凝集活性显著。使用甘露聚糖-Sepharose 4B亲和层析柱纯化菌株gs1总蛋白,得到凝集素。Brandford法定量检测分析表明,1000 ml gs1发酵培养液中含有9.58 mg 凝集素。SDS-PAGE 电泳分析显示该凝集素为单一条带,分子量约为12 kDa。凝集活性实验表明,该凝集素对兔、大鼠和小鼠的血红细胞具有凝集作用,对兔血红细胞效果最显著;而对人(A\B\O\AB型)和鸡的血红细胞无凝集作用。糖结合活性实验表明,甘露糖对该凝集素的凝集活性具有抑制作用。通过初步分类鉴定,菌株gs1为半知菌亚门,丝孢纲,丛梗孢目,丛梗孢科,曲霉属。 相似文献
4.
Platelet plasma membrane lectin activity 总被引:5,自引:0,他引:5
T K Gartner D C Williams D R Phillips 《Biochemical and biophysical research communications》1977,79(2):592-599
The lectin activity of human platelet and erythrocyte membranes was evaluated using trypsinized, formalinized erythrocytes from eight species. Platelet membranes had the greatest lectin activity against cow erythrocytes, but also had significant activity against human, sheep, electric eel, and rabbit erythrocytes. In contrast, erythrocyte membranes only had low lectin activity against electric eel erythrocytes with no activity against the other types of erythrocytes tested. The platelet membrane lectin activity was found to reside in protein molecules on the external surface of the platelet plasma membrane. The lectin activity of platelet membranes was inhibited by amino sugars and some basic amino acids: N-acetylated amino sugars and other neutral sugars were without effect. These results demonstrate that the external surface of the platelet plasma membrane has a specific lectin activity. 相似文献
5.
Related mannose-specific lectins from different species of the family Amaryllidaceae 总被引:5,自引:0,他引:5
Bulbs from three species of the plant family Amaryllidaceae ( Narcissus pseudonurcissus L., Leucojum aestivum L. and Leucojum vernum L.) were found to contain mannose-specific lectins. These lectins were serologically identical to a previously reported Amaryllidaceae lectin from Galanthus nivalis L. bulbs, but had a different molecular structure. The lectins described in this paper are dimeric proteins composed of subunits of 13 kDa, which are not held together by disulphide bridges. In hapten-inhibition assays Amaryllidaceae lectins exhibited exclusive specificity towards mannose. Furthermore, they all had a high specific agglutination activity with trypsin-treated rabbit erythrocytes, whereas human red blood cells were not agglutinated. 相似文献
6.
Ram Sarup Singh Ranjeeta Bhari Jatinder Singh Ashok Kumar Tiwary 《World journal of microbiology & biotechnology》2011,27(3):547-554
Mucin-specific lectin from mycelium of Aspergillus nidulans was purified using anion exchange and gel filtration chromatographic techniques with an overall recovery of 32% and 21.97-fold
purification. The purified lectin migrated as a single band in SDS–PAGE with an apparent molecular mass of 34 kDa. Carbohydrate
analysis revealed that it is a glycoprotein with total sugar content of 2.54%. Optimal agglutination was observed when serially
diluted lectin was incubated with human type O erythrocyte suspension at pH 7.0–8.0 and temperature 20–30°C. Lectin was found
to be completely stable within pH 5.0–8.0 and temperature at or below 40°C. Demetallization by extensive dialysis against
EDTA did not alter its haemagglutination activity. Lectin activity was reduced to half after 24 h incubation with urea and
thiourea, with no such effect of guanidine HCl. The lectin showed potent mitogenic response towards mouse splenocytes, attaining
a maximum at 200 μg/ml as compared to untreated control cells. Mitogenic lectins are invaluable tools to assess the functioning
of immune cells. None of the microfungal lectin has yet been investigated for mitogenic activity. This is the first report
on mitogenic activity of lectin from Aspergillus sp. 相似文献
7.
Zenteno R Vázquez L Martínez-Cairo S Bouquelet S Agundis C Zenteno E 《Glycoconjugate journal》2000,17(5):339-347
From the serum of juvenile freshwater prawns, we isolated by affinity chromatography on glutaraldehyde-fixed rat erythrocytes stroma, immobilized in Sephadex G-25, a sialic acid specific lectin of 9.6[emsp4 ]kDa per subunit. Comparative analysis against adult organisms purified lectin, by chromatofocusing, showed that the lectin from juvenile specimens is composed by four main isoforms with a pl of 4.2, 4.6, 5.1, and 5.6, whereas the lectin from adults is eluted at pH 4.2. The amino acid composition of the lectin obtained from adult and juvenile stages suggest identity, but the compositions are not identical since a higher content of carbohydrates was found in the lectin from younger organisms. The freshwater prawn lectin showed specificity toward N-acetylated amino sugar residues such as GlcNAc, GalNAc, Neu5Ac and Neu5,9Ac; but in juvenile organisms the lectin showed three times less hemagglutinating activity than the lectin from adults. Both lectins agglutinated rat, rabbit and chicken erythrocytes, indicating that Neu5,9Ac in specific O-glycosydically linked glycans seems to be relevant for the interaction of M. rosenbergii lectins with their specific cellular receptor. Our results suggest that the physicochemical characteristics of the lectin from the freshwater prawn are regulated through maturation. 相似文献
8.
Investigation on the occurrence of soluble lectins in mammalian nervous tissue extracts 总被引:3,自引:0,他引:3
R Joubert M Caron D Bladier 《Comparative biochemistry and physiology. B, Comparative biochemistry》1986,85(4):859-863
Five brain or retina crude extracts obtained from adult mammalians and nine fractions of brain extracts prepared by chromatography were screened for their lectin activities. All crude extracts and several fractions contained agglutinins reacting with neuraminidase-treated rabbit red blood cells. Hemagglutination activity varied widely with the method of preparation of the extracts. Hemagglutination inhibition tests were carried out to look for possible differences in the specificities of the agglutinins. All were found to be D-galactosyl specific. Each crude extract was found to contain a second lectin activity, which was detected using ethanol-treated rabbit erythrocytes known to react with heparin-binding lectins. Hemagglutination and inhibition studies showed that they completely differ from the galactoside-binding lectins detected previously. The possible functions of these lectins are discussed. 相似文献
9.
Along with a brief review of Lentinula edodes (shiitake mushroom) submerged cultivation history within the framework of important extracellular proteins biosynthesis,
this study contains the authors’ own results. The possibility of regulating the lectin activity of shiitake using the synthetic
components is shown. The time course of lectin production in culture liquid of L. edodes in different media under submerged culture conditions was studied. The activity of agglutinins depended on the ratio between
carbon and nitrogen sources and the pH of the culture medium. A relationship between the chemical composition of nutrient
medium, the activity of extracellular lectins of L. edodes, and the formation of pigmented mycelial film in liquid culture has been found. The formulation of medium, on which the brown
mycelial film appears in several days of submerged cultivation, is proposed. The results obtained make a contribution to the
present notion of biochemical processes that give rise to the occurrence of the aforesaid morphological structure of shiitake.
Finally, two extracellular lectins from the submerged culture of L. edodes have been isolated and purified to homogeneity. Their physicochemical properties and composition have been studied. 相似文献
10.
Hamshou M Smagghe G Van Damme EJ 《Communications in agricultural and applied biological sciences》2007,72(3):639-644
Lectins are carbohydrate-binding proteins that contain at least one carbohydrate binding domain which can bind to a specific mono- or oligosaccharide. These proteins are widely distributed in plants. However, over the last decade evidence is accumulating that lectins occur also in numerous fungi belonging to both the Ascomycota and Basiodiomycota. Rhizoctonia solani is known to be an important pathogen to a wide range of host plants. In this study, isolates of R. solani from different anastomosis groups have been screened for the presence of lectin using agglutination assays to detect and quantitate lectin activity. The evaluation included determination of the lectin content in mycelium as well as in sclerotia. The amount of lectin in the sclerotia was higher than in the mycelium of the same strains. The R. solani strains with the highest amounts of lectin have been selected for cultivation, extraction and purification of the lectin. 相似文献
11.
Along with a brief review of Lentinula edodes (shiitake mushroom) submerged cultivation history within the framework of important extracellular proteins biosynthesis, this study contains the authors’ own results. The possibility of regulating the lectin activity of shiitake using the synthetic components is shown. The time course of lectin production in culture liquid of L. edodes in different media under submerged culture conditions was studied. The activity of agglutinins depended on the ratio between carbon and nitrogen sources and the pH of the culture medium. A relationship between the chemical composition of nutrient medium, the activity of extracellular lectins of L. edodes, and the formation of pigmented mycelial film in liquid culture has been found. The formulation of medium, on which the brown mycelial film appears in several days of submerged cultivation, is proposed. The results obtained make a contribution to the present notion of biochemical processes that give rise to the occurrence of the aforesaid morphological structure of shiitake. Finally, two extracellular lectins from the submerged culture of L. edodes have been isolated and purified to homogeneity. Their physicochemical properties and composition have been studied. 相似文献
12.
Lectins from seeds of nine species of Erythrina have been purified by affinity chromatography on columns of lactose coupled to Sepharose and their properties compared with those of the lectin from Erythrina cristagalli. All lectins are glycoproteins of M, ca 60 000 composed of two identical or nearly identical subunits. They contain between 3–10% carbohydrates comprised of N-acetylglucosamine, mannose, fucose and xylose. The amino acid composition of all Erythrina lectins is very similar. The N-terminal amino acid is valine, with the exception of the lectin from E. flabelliformis in which it is alanine. To the extent tested, identities or near identities have been found in the N-terminal sequences (up to 15 residues in some cases) of the lectins. Hapten inhibition experiments of agglutination have shown that the lectins are specific for N-acetyllactosamine, this disaccharide being 10–30 times more inhibitory than D-galactose and 10–20 times more than N-acetyl-D-galactosamine. All lectins agglutinate human erythrocytes equally well, irrespective of blood type, at minimal concentrations of 5–20 μg/ml. Six of the lectins are also very effective in agglutinating rabbit erythrocytes and are mitogenic for human peripheral blood lymphocytes, whereas three of them are considerably weaker hemagglutinins for rabbit erythrocytes, and two of these are also very weak mitogens. Our results, while demonstrating striking similarities in the molecular properties and sugar specificity of all Erythrina lectins studied, suggest the existence of differences at or close to the carbohydrate-binding site. 相似文献
13.
Matteo Cammarata Gigliola Benenati Eric W. Odom Giuseppina Salerno Aiti Vizzini Gerardo R. Vasta Nicolò Parrinello 《Biochimica et Biophysica Acta (BBA)/General Subjects》2007
A novel fucose-binding lectin, designated SauFBP32, was purified by affinity chromatography on fucose–agarose, from the serum of the gilt head bream Sparus aurata. Electrophoretic mobility of the subunit revealed apparent molecular weights of 35 and 30 kDa under reducing and non-reducing conditions, respectively. Size exclusion analysis suggests that the native lectin is a monomer under the selected experimental conditions. Agglutinating activity towards rabbit erythrocytes was not significantly modified by addition of calcium or EDTA; activity was optimal at 37 °C, retained partial activity by treatment at 70 °C, and was fully inactivated at 90 °C. On western blot analysis, SauFBP showed intense cross-reactivity with antibodies specific for a sea bass (Dicentrarchus labrax) fucose-binding lectin. In addition, the similarity of the N-terminal sequence and a partial coding domain to teleost F-type lectins suggests that SauFBP32 is a member of this emerging family of lectins. 相似文献
14.
Phagocytic activity of three Naegleria strains in the presence of erythrocytes of various types 总被引:1,自引:0,他引:1
The phagocytic activities of N. lovaniensis (Aq/9/1/45D) and N. gruberi (1518/1f and 1518/1e) were studied in the presence of erythrocytes of various species: chicken, rabbit, goat, and human (A+, B+, and AB+ were tested). The percentage of amoebae with ingested red cells, the phagocytic index (PhI), can be considered as an expression of phagocytic activity. Under given conditions (erythrocyte concentration, incubation time, age of amoebic cultures) each strain of Naegleria prefers one erythrocyte type. Thus, for 72-h cultures, N. lovaniensis ingested more A+ type erythrocytes than did N. gruberi strains but had very low affinity for rabbit red cells except when very high concentrations were tested. Naegleria gruberi 1f was the most active of the three strains towards rabbit and B+ and AB+ human erythrocytes, but very low PhIs were obtained with goat erythrocytes. Naegleria gruberi 1e exhibited high phagocytic activity for every erythrocyte type except for rabbit red cells. 相似文献
15.
Blackgram (Vigna mungo
L. Hepper)seeds contain two galactose-specific lectins, BGL-I and BGL-II. BGL-I was partially purified into two monomeric lectins which
were designated as BGL-I-1 (94 kDa) and BGL-I-2 (89 kDa). BGL-II is a monomeric lectin of 83 kDA. The purified lectins were
associated with galactosidase activities. BGL-I-1 and BGL-II were copurified with α-galactosidase activity while BGL-I-2 was
largely associated with β-galactosidase activity. These lectins agglutinate trypsin treated rabbit erythrocytes, but not the
human erythrocytes of A, B or O groups. They were stable between pH 3·5 and 7·5 for their agglutination. The lectins did not
show any metalion requirement. They were inactivated at 50°C. The lectin activity was inhibited by D-galactose (0·1 mM). The
Scatchard plots of galactose binding to these lectins are nonlinear and biphasic curves indicative of multiple binding sites.
The data show that the monomeric lectins have both lectin and galactosidase activities suggestive of a bifunctional protein. 相似文献
16.
Qiuxiang Pang Xuemei Liu Bosheng Zhao Wei Wei Xiufang Zhang Lianfei Zhao Jingjing Xie Huanhuan Sun 《Central European Journal of Biology》2012,7(2):354-361
Lectins are important components of the immune defense system of invertebrates. Given their important functions, numerous
investigations have been carried out on the characterization and function of lectins in invertebrates. However, lectin studies
with the freshwater planarian, an evolutionarily important animal, are rare. In this paper, we demonstrate agglutination of
glutaraldehyde treated erythrocytes by a lectin with preference for rabbit erythrocytes. The result of hemagglutinating activity
inhibition assays with several carbohydrates showed the most potent inhibitor was maltose. A natural lectin from the crude
homogenates of freshwater planarian Dugesia japonica was purified by single step affinity chromatography using amylose-coupled agarose. The purified protein appeared as one band
with a molecular mass of 350 kDa in PAGE, and as one band, approximately 56 kDa, in SDS-PAGE. The purified lectin showed dependence
on calcium. The activity of the purified lectin was inhibited at temperatures greater than 50°C and showed a pH optimum between
5–8. The purified lectin also has binding activity to the Gram-negative bacteria E. coli, and the Gram-positive bacteria B. subtilis. Furthermore, the purified lectin obtained from injured and bacteria-induced planarians showed increased agglutinating activity
against rabbit erythrocytes. These results suggest that the purified lectin may play an important role in the innate immunity
of the freshwater planarian. 相似文献
17.
A number of lectins varying in polypeptide composition and carbohydrate specificity were isolated from Lentinus edodes at different stages of its morphogenesis: nonpigmented mycelium, brown mycelium film, and fruiting body. Three lectins were identified at the nonpigmented mycelium stage, two of them being dimers consisting of 16 and 45 kDa and 16 and 42 kDa subunits; the third is a tetramer of 16, 39, 42, and 45 kDa subunits. The fractions with lectin activity obtained at the brown mycelium film stage contained polypeptides of 24, 30, and 38 kDa, characteristic of this morphological structure. The fruiting body was shown to contain two lectins of 43 and 55 kDa. All of the isolated lectins expressed the highest affinity towards L,D-melibiose, D-lactose, and D-galactose. 相似文献
18.
The nature of the receptors for T1 and T4 Neisseria gonorrhoeae on erythrocytes and other cells was investigated. In general, cells of nonprimate origin contained few receptors for gonococci. Receptors for T4 gonococci were only uncovered when host cells were pretreated with trypsin. Trypsinization, while unnecessary for T1 adherence to erythrocytes, enhanced attachment in inverse proportion to original erythrocyte sensitivity. Receptors for T1 and T4 organisms on trypsinized and trypsin-neuraminidase-treated erythrocytes were blocked by concanavalin A and peanut lectins, respectively, but a distinction could be made between them with wheat germ lectin and galactose oxidase. Of a number of sugars tested as inhibitors, only D-galactose blocked adherence of T4 but was without effect on T1. While the identity of erythrocyte receptors is uncertain, likely candidates are "band 3" protein and glycophorin, by virtue of their galactose content, lectin binding capacity, and partial exposure on the outer surface of the erythrocyte. 相似文献
19.
Normal bovine erythrocytes were agglutinated with four of five lectins specific for different oligosaccharides. The order of reactivity was wheat germ greater than ricin greater than soybean greater than peanut. Concanavalin A did not agglutinate normal bovine erythrocytes. After neuraminidase treatment of normal bovine erythrocytes, each lectin agglutinated the cells with decreased concentrations of lectin, verifying that partial removal of sialic acid exposes more of each lectin's binding sites or alters the binding site such that fewer molecules of lectin are required to initiate agglutination. A change in agglutination of erythrocytes using soybean agglutinin and peanut agglutinin occurred when cells were obtained from cattle infected with Eperythrozoon wenyoni. The results suggested that an alteration in erythrocyte membranes occurred as a result of this infection as manifested by the increased recognition of both the soybean agglutinin and peanut agglutinin receptor carbohydrates. A similar effect was indicated with erythrocytes obtained during an acute Anaplasma marginale infection; however, an ensuing reticulocytosis masked the effect, requiring the use of fluoresceinated lectins to verify that increased binding of each lectin occurred with infected cells when compared to normal cells. 相似文献
20.
Singh J Singh J Kamboj SS 《Biochemical and biophysical research communications》2004,318(4):1057-1065
A novel lectin having specificity towards a complex glycoprotein asialofetuin was purified from tubers of Arisaema flavum (Schott.) by affinity chromatography on asialofetuin-linked amino-activated silica beads. A. flavum gave a single peak on HPLC size exclusion and a single band on non-denatured PAGE at pH 4.5. The molecular mass of the lectin, as determined by gel filtration chromatography, was 56 kDa. In SDS-PAGE, pH 8.3, the lectin migrated as a single band of 13.5 kDa, under reducing and non-reducing conditions, indicating the homotetrameric nature. A. flavum lectin (AFL) readily agglutinated rabbit, rat, sheep, goat, and guinea pig erythrocytes but not human ABO blood group erythrocytes even after neuraminidase treatment. This lectin is stable up to 55 degrees C and does not require metal ions for its hemagglutination activity. AFL was completely devoid of sulphur containing amino acids and was rich in aspartic acid and glycine. In Oucterlony's double immunodiffusion, the antisera raised against A. flavum lectin showed distinct lines of identity with those of other araceous lectins. AFL showed potent mitogenic activity towards BALB/c splenocytes and human lymphocytes in comparison to Con A, a well-known plant mitogen. AFL also showed significant in vitro antiproliferative activity towards J774 and P388D1 murine cancer cell lines. 相似文献