scholarly journals Membrane glycoproteins of the nerve growth cone: diversity and growth regulation of oligosaccharides.

1986 ◽  
Vol 103 (4) ◽  
pp. 1369-1382 ◽  
Author(s):  
L M Greenberger ◽  
K H Pfenninger
Keyword(s):  
Sialic Acid ◽  
Growth Cone ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Growth Regulation ◽  
Binding Proteins ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Adult Brain ◽  
Two Dimensional

A subcellular fraction prepared from fetal rat brain and enriched in growth cone membranes is analyzed for its lectin-binding proteins. Growth-associated glycoproteins are identified by comparing the growth cone glycoproteins with those of synaptosomes. Protein was resolved in one- or two-dimensional gels, electroblotted, and blots probed with radioiodinated concanavalin A, wheat germ agglutinin, and Ricinus communis agglutinins I and II. In one-dimensional gels, each lectin recognizes approximately 20 polypeptides (with substantial overlap) most of which migrate diffusely and have relatively high molecular masses (range 30-200 kD). The seven major Coomassie-staining proteins of the membrane fraction (34-52 kD) are not the major lectin-binding proteins. In two-dimensional gels, the lectin-binding proteins are either streaked across the pH gradient or exist as multiple spots, indicating broad charge heterogeneity. Seven wheat germ agglutinin- and Ricinus communis agglutinin II-binding glycoproteins are present in greater abundance in growth cone fractions compared with synaptosomes. Most notably, an acidic, sialic acid-rich protein (27-30 kD, pI 4.0; termed gp27-30) is most abundant at postnatal day 4, but absent from adult brain. The protein's very acidic isoelectric point is due, at least in part, to its high sialic acid content. Growth regulation of specific protein-linked oligosaccharides suggests that they play a special role in growth cone function. In addition, the great diversity of growth cone glycoproteins from whole brain suggests glycoprotein heterogeneity among growth cones from different neuron types.

Changing surface antigen and carbohydrate patterns during the development of Oesophagostomum dentatum

Parasitology ◽  
1999 ◽  
Vol 119 (5) ◽  
pp. 491-501 ◽  
Author(s):  
A. JOACHIM ◽  
B. RUTTKOWSKI ◽  
A. DAUGSCHIES
Keyword(s):  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Soybean Agglutinin ◽  
Peanut Agglutinin ◽  
Oesophagostomum Dentatum ◽  
Periodate Treatment

Living and fixed specimen of Oesophagostomum dentatum were labelled in situ with serum antibodies or a panel of biotin- labelled lectins. Specific binding of antibodies was observed in all parasitic stages – freshly exsheathed 3rd-stage larvae (L3), 3rd- and 4th-stage (L4) larvae cultured in vitro and L3 and L4 and adults isolated from pig intestines. The shedding of the stained layer by motile larvae was inhibited by levamisole-induced paralysis. Larvae cultured in vitro exposed serum-derived proteins on their surface which could be labelled with secondary antibody directed against the respective serum donor species. While freshly exsheathed larvae were recognized by O. dentatum-positive serum only, older larvae and adults cross-reacted with serum from pigs infected with O. quadrispinulatum, a closely related species. Lectin binding varied considerably between stages. While binding was not observed in pre-parasitic stages, Concanavalin A, Soybean Agglutinin, Wheat Germ Agglutinin, Ricinus communis Agglutinin and Peanut Agglutinin bound to developing larvae in varying degrees. Dolichos biflorus Agglutinin only bound to advanced (luminal) larval stages, while adults generally displayed only weak or partial lectin binding (except with Concanavalin A and Wheat Germ Agglutinin). Ulex europaeus Agglutinin only labelled larvae derived from cultures containing 10% pig serum. Cleavage of the carbohydrate residues by sodium periodate treatment resulted in reduction of antibody binding to cultured larvae, but not to freshly exsheathed L3. Concanavalin A, Soybean Agglutinin, and Peanut Agglutinin binding was also reduced by periodate treatment, while binding of Wheat Germ Agglutinin and Ricinus communis Agglutinin was inhibited only in early L3, but not in older stages. The different lectin labelling patterns are related to the different stages of the nematode – infective, invasive, histotropic, and luminal – and may serve as a mode of adaptation for the parasite against the host's immune attack by surface glycoprotein variation, together with antigen shedding (as demonstrated by labelling of motile larvae) and a possible acquisition of host molecules at the parasite's surface. Furthermore, a possible role of this developmental variation in surface carbohydrates in parasite–parasite interactions is discussed.

scholarly journals A cytochemical study of lectin receptors on isolated chick neural retina neurons in vitro

1982 ◽  
Vol 53 (1) ◽  
pp. 1-20
Author(s):  
J.A. Bee
Keyword(s):  
Sialic Acid ◽  
Cell Body ◽  
Growth Cone ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Neuronal Cell ◽  
Cytochemical Study ◽  
Lectin Receptors ◽  
Dolichos Biflorus ◽  
Dolichos Biflorus Agglutinin

The cell body, neurite and growth cone of isolated retinal neurons have been compared on the basis of their ability to bind a number of fluorescently labelled lectins, each possessing a unique carbohydrate specificity. The susceptibility of the respective binding patterns following pretreatment of these fixed cells with either neuraminidase or trypsin was also investigated. Neuronal cell bodies displayed the most intense binding of each lectin, with localization of limulin binding (specific for sialic acid) predominantly to the neurite hillock, the point on the cell body from which the neurite projects. Limulin binding was almost totally abolished by pretreatment with either neuraminidase or trypsin. In contrast to the cell body, limulin binding to the neurite or growth cone was not detected. These regions of the cell apparently possessed sialic acid, however, since pretreatment with neuraminidase reduced wheat germ agglutinin binding (to N-acetylglucosamine) and markedly enhanced Dolichos biflorus agglutinin binding (to N-acetylgalactosamine) to both the neurite and growth cone. The initially low binding of Dolichos biflorus agglutinin to the neurite and growth cone was slightly enhanced by pretreatment with trypsin. Uniformly low levels of binding of either Ricinus communis agglutinin 60 (galactose, N-acetylgalactosamine) or R. communis agglutinin 120 (galactose) was observed over the entire neuron. R. communis agglutinin 120 binding was not enhanced by pretreatment with neuraminidase. Receptors for either concanavalin A (mannose, glucose) or Ulex europaeus agglutinin I (fucose) were abundant over the entire nerve cell with the former exhibiting more marked trypsin sensitivity. From these data, it is apparent that the repertoire of lectin binding sites of the neurite and growth cone of these differentiating nerve cells differs markedly from that of the cell body, which itself demonstrates some degree of regionalization.

Prespore and prestalk differentiation of Dictyostelium discoideum as examined by changes of wheat germ agglutinin binding proteins

1987 ◽  
Vol 65 (1) ◽  
pp. 68-75 ◽  
Author(s):  
Akiko Kumagai ◽  
Hideo Mori ◽  
Shinoi Osuka ◽  
Koji Okamoto
Keyword(s):  
Dictyostelium Discoideum ◽  
Wheat Germ Agglutinin ◽  
Culture System ◽  
Wheat Germ ◽  
Binding Proteins ◽  
Normal Development ◽  
Maximum Level ◽  
Cell Types ◽  
The Other ◽  
Two Dimensional

Prespore- and prestalk-specific, wheat germ agglutinin (WGA) binding proteins in Dictyostelium discoideum were identified on two-dimensional gels by the use of a peroxidase–antiperoxidase method. Using these proteins as markers, differentiation of the two presumptive cell types was examined during the development. In normal development, two groups of prespore-specific WGA-binding proteins were found: one was detectable when cells formed discrete aggregates without tips (10.5 h) and reached a maximum level at 12 h, while the other appeared at the time of slug formation (17 h). On the other hand, a prestalk-specific WGA-binding protein, having an unusually high pI value (pI ca. 9.5), began to accumulate just before slug formation (13.5 h). The changes of WGA-binding proteins in a shake culture system were similarly analysed and compared with those in normal development.

scholarly journals Isolation and partial characterization of the major sialoglycoprotein of human T-lymphoblastoid cells of a MOLT-4B cell line

1978 ◽  
Vol 175 (3) ◽  
pp. 823-831 ◽  
Author(s):  
M Saito ◽  
S Toyoshima ◽  
T Osawa
Keyword(s):  
Sialic Acid ◽  
Cell Line ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Exchange Chromatography ◽  
Total Carbohydrate ◽  
Lymphoblastoid Cells ◽  
Sugar Chain ◽  
Sugar Chains

A sialoglycoprotein with an approx. mol.wt. of 95000 was isolated from human lymphoblastoid cells of a MOLT-4B cell line, which was of human T-lymphocyte origin, by ion-exchange chromatography, affinity chromatography on a column of wheat-germ agglutinin-Sepharose and preparative slab-gel electrophoresis. The localization of this glycoprotein on the cell surface was indicated by surface labelling by the periodate/NaB3H4 and lactoperoxidase-catalysed iodination methods. Carbohydrate analyses of this glycoprotein revealed that its total carbohydrate content is 28% (w/w), and it contains fucose, galactose, mannose, N-acetylglucosamine, N-acetylgalactosamine and sialic acid in molar proportions 1.0:4.0:3.7:3.5:1.2:2.5, suggesting that it has two types of sugar chain, i.e. sugar chains like those of serum glycoproteins and sugar chains of the type found in mucins. Actually, alkaline borohydride treatment of this glycoprotein yielded tri- and tetra-saccharide, the latter containing 1 molecule of fucose in addition to each molecule of galactose, N-acetylgalactosamine and sialic acid. This glycoprotein bound to Ricinus communis agglutinin and concanavalin A as well as to wheat-germ agglutinin.

Lack of binding of serum glycoprotein hormone α subunit to Concanavalin A–Sepharose reflects increased branching of the oligosaccharide chains

1984 ◽  
Vol 103 (1) ◽  
pp. 111-116 ◽  
Author(s):  
A. J. Chapman ◽  
J. T. Gallagher ◽  
C. G. Beardwell ◽  
S. M. Shalet
Keyword(s):  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Pituitary Tumours ◽  
Con A ◽  
Glycoprotein Hormone ◽  
Α Subunit ◽  
Binding Forms

ABSTRACT The lectin-binding properties of serum α subunit were studied by lectin affinity chromatography. Normal individuals and most patients with pituitary tumours produced α subunit which bound specifically to Concanavalin A–Sepharose (Con A). Some patients with pituitary tumours produced both Con A-reactive α subunit and α subunit which did not bind to Con A. Concanavalin A–Sepharose-binding α subunit from all sources bound strongly to Ricinus communis agglutinin–Sepharose after treatment with neuraminidase. Serum α subunit from those patients with pituitary tumours, which did not bind to Con A, bound to wheat germ agglutinin–Sepharose, exhibiting both weakly binding and strongly binding forms. Serum α subunit from both patients and controls, which did bind to Con A, showed only weak affinity for wheat germ agglutinin–Sepharose. Neither the low affinity nor the high affinity of serum α subunit from any source for wheat germ agglutinin–Sepharose was affected by neuraminidase. These findings show that (a) the predominant pattern of glycosylation of serum α subunit from normal controls is a Con A-reactive, biantennate complex oligosaccharide and (b) that the structural alteration which results in serum α subunit which does not bind to Con A in some patients with pituitary tumours is not an absence of carbohydrate, rather the α subunit contains highly branched, either complex or hybrid oligosaccharides. J. Endocr. (1984) 103, 111–116

scholarly journals Sialic acid in mature megakaryocytes: detection by wheat germ agglutinin

Blood ◽  
1985 ◽  
Vol 65 (5) ◽  
pp. 1120-1126
Author(s):  
PK Schick ◽  
WG Jr Filmyer
Keyword(s):  
Sialic Acid ◽  
Guinea Pig ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Lectin Binding ◽  
Fluorescence Emission ◽  
Stage Iv ◽  
Computer Assisted ◽  
Chromomycin A3 ◽  
Vibrio Cholera

The characteristics of the surface of guinea pig megakaryocytes were investigated with wheat germ agglutinin (WGA). Purified guinea pig megakaryocytes and platelets were incubated with WGA conjugated with rhodamine, cytocentrifuged, and then exposed to Chromomycin A3 for the assessment of ploidy. The fluorescence emission of the DNA-Chromomycin complex was similar to that of fluorescein, and thus both rhodamine-WGA and Chromomycin A3 fluorescence could be analyzed in the same cell. Ploidy was assessed by microdensitometry of Chromomycin A3 fluorescence. Eight hundred megakaryocytes were analyzed by four parameters: (1) labeling by WGA, (2) ploidy, (3) morphological stage, and (4) size. The results were analyzed by a computer-assisted program. Although all platelets had reacted with WGA, only about half of the isolated megakaryocytes had been labeled by the lectin. The analysis of the megakaryocytes that had reacted revealed that 72% of stage III and 77% of stage IV megakaryocytes as compared with 35% of stage I and 29% of stage II cells had been labeled by the lectin. WGA reacted with 44% of 8N megakaryocytes and 60% and 59% of 16N and 32N cells. However, WGA labeling was independent of megakaryocyte size. The digestion of 15% and 48% of megakaryocyte sialic acid with neuraminidase from Vibrio cholera resulted in a 67% and 89% decrease in the binding of rhodamine- WGA, respectively, as determined by microdensitometry. The study indicated that sialic acid serves as a receptor for WGA and that sialoglycoproteins and possibly gangliosides become exposed on the surface of mature megakaryocytes. WGA can recognize mature megakaryocytes by biochemical criteria and the assessment of lectin binding could complement the morphological staging of megakaryocytes.

scholarly journals Lectin receptors as markers for Trypanosoma cruzi. Developmental stages and a study of the interaction of wheat germ agglutinin with sialic acid residues on epimastigote cells.

1980 ◽  
Vol 152 (5) ◽  
pp. 1375-1392 ◽  
Author(s):  
M E Pereira ◽  
M A Loures ◽  
F Villalta ◽  
A F Andrade
Keyword(s):  
Sialic Acid ◽  
Cell Surface ◽  
Trypanosoma Cruzi ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Lectin Binding ◽  
The Other ◽  
Con A ◽  
Bauhinia Purpurea ◽  
Binding Lectin

Trypanosoma cruzi at various stages of maturation and differentiation have been isolated by conventional cellular fractionation procedures and characterized by cell surface markers using 30 highly purified lectins encompassing all known sugar specificities. Cell surface carbohydrates of the various T. cruzi stages were analyzed by agglutination and lectin-binding assays. Specific receptors for wheat germ agglutinin (WGA), Helix pomatia, Sophora japonica, and Bandeiraea simplicifolia lectin II were found only in culture epimastigotes, whereas peanut agglutinin (PNA) sites were present exclusively in amastigotes, those for Phaseolus vulgaris in bloodstream trypomastigotes and amastigotes, and for Wistaria floribunda hemagglutinin predominantly in culture forms of T. cruzi. The N-acetylgalactosamine (DGalNAc)-binding lectin from Bauhinia purpurea agglutinated and inhibited the movement of epimastigotes and bloodstream trypomastigotes, but it only inhibited--without agglutinating--culture trypomastigotes. Because both the agglutination and inhibition of movement were reversed by specific sugar haptens, Bauhinia purpurea sites were present in all the flagellated parasites. On the other hand, PNA sites were detectable on epimastigotes after the cells were treated with sialidase, whereas, at the same time, WGA receptors were completely removed and those for the other sialic acid-binding proteins, Aaptos papillata lectin II and Limulus polyphemus, were partially eliminated; moreover, the activity of Wistaria floribunda hemagglutinin, a DGalNAc-binding lectin, increased 4,000 times. Trypsinization and lyzozyme treatment of epimastigote cells did not significantly affect lectin agglutination or lectin binding. WGA reacted solely with sialic acid residues on epimastigote cell surface with an apparent association constant of 2 x 10(6) M-1, each epimastigote having an estimated average of 3 x 10(6) WGA sites, as determined by binding experiments and a minimum of 7.7 x 10(6) sialic acid residues, as calculated by colorimetric method after sialidase digestion. Evidences are presented that the sialyl residues are rapidly regenerated (in approximately 4 h) and that they, at least for the most part, are not adsorbed from the culture medium. The receptor for the D-mannose-binding lectins (concanavalin A [Con A] and Lens culinaris) must either be on the same carbohydrate moiety having the WGA site, or, if in a distinct molecule, both carrier molecules of Con A and WGA sites must be located close to each other in the plasma membrane of the parasite.

scholarly journals Sialic acid in mature megakaryocytes: detection by wheat germ agglutinin

Blood ◽  
1985 ◽  
Vol 65 (5) ◽  
pp. 1120-1126 ◽  
Author(s):  
PK Schick ◽  
WG Jr Filmyer
Keyword(s):  
Sialic Acid ◽  
Guinea Pig ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Lectin Binding ◽  
Fluorescence Emission ◽  
Stage Iv ◽  
Computer Assisted ◽  
Chromomycin A3 ◽  
Vibrio Cholera

Abstract The characteristics of the surface of guinea pig megakaryocytes were investigated with wheat germ agglutinin (WGA). Purified guinea pig megakaryocytes and platelets were incubated with WGA conjugated with rhodamine, cytocentrifuged, and then exposed to Chromomycin A3 for the assessment of ploidy. The fluorescence emission of the DNA-Chromomycin complex was similar to that of fluorescein, and thus both rhodamine-WGA and Chromomycin A3 fluorescence could be analyzed in the same cell. Ploidy was assessed by microdensitometry of Chromomycin A3 fluorescence. Eight hundred megakaryocytes were analyzed by four parameters: (1) labeling by WGA, (2) ploidy, (3) morphological stage, and (4) size. The results were analyzed by a computer-assisted program. Although all platelets had reacted with WGA, only about half of the isolated megakaryocytes had been labeled by the lectin. The analysis of the megakaryocytes that had reacted revealed that 72% of stage III and 77% of stage IV megakaryocytes as compared with 35% of stage I and 29% of stage II cells had been labeled by the lectin. WGA reacted with 44% of 8N megakaryocytes and 60% and 59% of 16N and 32N cells. However, WGA labeling was independent of megakaryocyte size. The digestion of 15% and 48% of megakaryocyte sialic acid with neuraminidase from Vibrio cholera resulted in a 67% and 89% decrease in the binding of rhodamine- WGA, respectively, as determined by microdensitometry. The study indicated that sialic acid serves as a receptor for WGA and that sialoglycoproteins and possibly gangliosides become exposed on the surface of mature megakaryocytes. WGA can recognize mature megakaryocytes by biochemical criteria and the assessment of lectin binding could complement the morphological staging of megakaryocytes.

scholarly journals The investigation of the changes in the surface glycoconjugates using two different spheroid models of breast cancer cells and availability assessment of these spheroid models for rapid diagnosis

2021 ◽  
Vol 38 (3) ◽  
pp. 266-271
Author(s):  
Yosun MATER ◽  
Günnur DEMİRCAN
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Sialic Acid ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Sialic Acids ◽  
Maackia Amurensis ◽  
Maackia Amurensis Lectin

The importance of early cancer diagnosis has led to development of many different diagnostic methods. In this context, the studies investigating the presence and amount of sugar residues to use as indicators in the identification of cancer cell type have become prominent. In the present study, sialic acids found on the membrane surfaces of ER (+) MCF-7 and ER (-) MDA-MB-231 breast cancer cell lines were labeled using three-dimensional (3D) cell culture (Spheroid) model as the closest method to the patient sample, thus its natural environment, among the cell culture methods. These sugar units that play a role in regulation of important immune characteristics such as recognition, binding and metastasis were made visualizable by applying fluorescent-labeled lectins such as FITC-(Wheat Germ Agglutinin) specifically binding to sialic acid units (GlcNAc, Neu5Ac) including particularly ß-GlcNAc and FITC-(Maackia Amurensis-Lectin-1) specifically binding to Galß4GlcNAc type sialic acids. These glycan units were specifically labeled with FITC-(Maackia Amurensis-Lectin-1) and FITC- (Wheat Germ Agglutinin) and radiation intensities were analyzed relatively. The two different breast cancer cell cultures were compared with respect to change in the amounts of sialic acid residues containing α-2,3- and α-2,6 bonds using fluorescent-labeled lectins. In the present study, we have performed a precise, accurate and rapid determination of the sugar content in the different breast cancer cell surface lines by means of fluorescent-labeled lectins and carried out a relative comparison between the micrographs.

Characterization of the exposed carbohydrates on the surface membrane of adult Schistosoma mansoni by analysis of lectin binding

Parasitology ◽  
1980 ◽  
Vol 81 (1) ◽  
pp. 1-15 ◽  
Author(s):  
A. J. G. Simpson ◽  
S. R. Smithers
Keyword(s):  
Schistosoma Mansoni ◽  
Adult Male ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Specific Binding ◽  
Lectin Binding ◽  
Surface Membrane ◽  
Soybean Agglutinin ◽  
Peanut Agglutinin

SUMMARYThe surface architecture of adult male Schistosoma mansoni was explored using a range of lectins with differing carbohydrate specificities. Highest specific binding was achieved with concanavalin A and the agglutinin of molecular weight 60000 from Ricinus communis; the binding of wheat germ agglutinin was mostly non-specific. Small amounts of peanut agglutinin and soybean agglutinin binding were observed and the binding of these lectins was increased by pre-treating the parasite with neuraminidase. The fucose binding protein of Lotus tetragonolobus failed to bind. These results indicate that mannose and/or glucose, galactose, N-acetyl-glucosamine, N-acetyl-galactosamine and sialic acid are exposed on the surface of the adult male schistosome.

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