Differential redistribution of lectin receptor classes on clonal rat myotubes and myoblasts

To evaluate the relative mobilities of cell surface glycoconjugates during myogenesis we have studied the redistribution of fluorescein-conjugated plant lectins on L6 rat myogenic cells. Previous experiments had demonstrated that the receptors for the lectins soybean agglutinin (SBA), wheat germ agglutinin, concanavalin A and Lens culinaris agglutinin all were relatively uniformly distributed on both myoblasts and myotubes, and that SBA receptors were capable of rapid redistribution on myotubes but not myoblasts at 4 degrees C (Sawyer & Akeson, 1983). Here we show that when SBA-labelled myoblasts are incubated at 37 degrees C, or for extended times at 4 degrees C, the lectin aggregates as on myotubes. So it appears that SBA-binding components show a quantitative rather than qualitative change in their mobility during L6 differentiation. In addition, the redistribution of the three other lectins on myoblasts and myotubes was either less prominent (i.e. showing fewer apparent surface clusters) or occurred less rapidly than with SBA. None of these three lectins showed striking differences in mobility between myoblasts and myotubes. Thus, it appears that SBA binds to a subset of surface glycoconjugates that is relatively highly mobile, and that this mobility is specifically enhanced with differentiation.

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Studies on the cell surface of zoospores and cysts of the fungus Phytophthora cinnamomi: nature of the surface saccharides as determined by quantitative lectin binding studies.

Journal of Histochemistry & Cytochemistry ◽

10.1177/33.5.3838761 ◽

1985 ◽

Vol 33 (5) ◽

pp. 384-388 ◽

Cited By ~ 16

Author(s):

A Bacic ◽

M L Williams ◽

A E Clarke

Keyword(s):

Cell Surface ◽

Concanavalin A ◽

Wheat Germ Agglutinin ◽

Wheat Germ ◽

Phytophthora Cinnamomi ◽

Lectin Binding ◽

Soybean Agglutinin ◽

Binding Studies ◽

Con A

The nature of the surface saccharides of zoospores, "partially encysted zoospores" and cysts of the root-rotting fungus Phytophthora cinnamomi, has been examined by quantitative lectin binding studies. Zoospores bound concanavalin A (Con A), but did not bind any of a variety of other lectins tested. In contrast, both cysts and "partially encysted zoospores" bound soybean agglutinin (SBA) as well as Con A. This indicates that accessible alpha-D-glucosyl/alpha-D-mannosyl-containing glycoconjugates predominate at the zoospore surface, whereas both alpha-D-glucosyl/alpha-D-mannosyl and galactosyl and/or N-acetyl-D-galactosaminosyl residues are accessible at the surface of cysts and "partially encysted zoospores." Neither Ulex europeus lectin nor wheat germ agglutinin (WGA) bound to any of the three cell preparations, indicating the absence of accessible alpha-L-fucosyl and N-acetyl-D-glucosaminosyl residues.

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Interaction of Lens culinaris lectin, Concanavalin A, Ricinus communis agglutinin and wheat germ agglutinin with the cell surface of normal and transformed rat liver cells

Experimentelle Pathologie ◽

10.1016/s0014-4908(75)80039-1 ◽

1975 ◽

Vol 10 (5-6) ◽

pp. 309-317 ◽

Cited By ~ 1

Author(s):

J. Roth ◽

G. Neupert ◽

K. Thoss

Keyword(s):

Cell Surface ◽

Rat Liver ◽

Concanavalin A ◽

Wheat Germ Agglutinin ◽

Wheat Germ ◽

Lens Culinaris ◽

Ricinus Communis ◽

Liver Cells ◽

Ricinus Communis Agglutinin ◽

Rat Liver Cells

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Factors Influencing the Reaction of α1-Fetoprotein with Concanavalin A and Lens Culinaris Agglutinin in Crossed Affinoimmunoelectrophoresis

Clinical Chemistry ◽

10.1093/clinchem/38.8.1418 ◽

1992 ◽

Vol 38 (8) ◽

pp. 1418-1424 ◽

Cited By ~ 5

Author(s):

D Magne ◽

N Seta ◽

D Lebrun ◽

G Durand ◽

D Durand

Keyword(s):

Concanavalin A ◽

Lens Culinaris ◽

Biological Fluids ◽

Con A ◽

Cellular Origin ◽

Lens Culinaris Agglutinin ◽

Lentil Lectin ◽

Wide Range ◽

Minimal Quantity ◽

Antibody Ratio

Abstract Concanavalin A (Con A) and lentil lectin (LCA) analysis of alpha-fetoprotein (AFP) glycosylation heterogeneity is used in a variety of clinical situations. We studied the influence of analytical conditions on the separation of AFP glycoforms by using lectin-crossed affinoimmunoelectrophoresis, regardless of the AFP concentration, which can vary over a wide range in biological fluids. We defined the optimal concentration of Con A (2 g/L) and LCA (0.35 g/L) in the first-dimension gel, together with the optimum antigen (AFP)/antibody ratio in the second-dimension gel. The presence of protein in the diluent used for AFP samples was found to change the shape of crossed affinoimmunoelectrophoresis patterns without changing the percentage composition of AFP fractions. The within-run CV was less than 4% for both lectins, and the between-run CV was less than 6.3%. The minimal quantity of AFP that provided a visible pattern with both lectins was 4 ng, corresponding to 50 microL of an 80 micrograms/L AFP sample. These technical conditions allow the cellular origin of AFP to be determined, regardless of the concentration in the sample. Typical AFP lectin patterns of secreting tumors are compared with fetal and cord serum AFP.

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Concanavalin A and Lens culinaris agglutinin binding patterns in normal and neoplastic sebaceous epithelium

Journal of Cutaneous Pathology ◽

10.1111/j.1600-0560.1986.tb01523.x ◽

1986 ◽

Vol 13 (4) ◽

pp. 274-284 ◽

Cited By ~ 1

Author(s):

L. C. Kuster ◽

B. H. Liwnicz ◽

D. E. Ward

Keyword(s):

Concanavalin A ◽

Lens Culinaris ◽

Lens Culinaris Agglutinin

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Concanavalin a and wheat germ agglutinin receptors on dictyostelium: Their visualization by scanning electron microscopy with microspheres

The Journal of Cell Biology ◽

10.1083/jcb.71.1.314 ◽

1976 ◽

Vol 71 (1) ◽

pp. 314-322 ◽

Cited By ~ 28

Author(s):

R Molday ◽

R Jaffe ◽

D McMahon

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Cell Surface ◽

Wheat Germ ◽

Cellular Interactions ◽

Con A ◽

Stages Of Development ◽

Lectin Receptors ◽

Plant Lectins ◽

Scanning Electron

The cellular slime mold, Dictyostelium discoideum, is a convenient model for studying cellular interactions during development. Evidence that specific cell surface components are involved in cellular interactions during its development has been obtained by Gerisch and co-workers (1, 2) using immunological techniques. Smart and Hynes (3) have shown that a cell surface protein can be iodinated on cells in aggregation phase, but not in vegetative phase, by the lactoperoxidase procedure. Recently, McMahon et al. (4), and Hoffman and McMahon have demonstrated, by SDS gel electrophoresis, considerable differences in cell surface proteins and glycoproteins of plasma membranes isolated from cells at different stages of development. Plant lectins have also been used to monitor changes in cell surface properties of D. discoideum cells during development. Weeks and co-workers (5, 6) have detected differences in the binding and agglutination of cells by concanavalin A (Con A). Gillette and Filosa (7) have shown that Con A inhibits cell aggregation and prematurely induces cyclic AMP phosphodiesterase. Capping of Con A receptors has also been reported (8). Reitherman et al. (9) have recently reported that agglutination of cells by several plant lectins and the slime mold agglutination, discoidin, changes during development. Such studies indicate that differences in surface properties exist for cells at various stages of development. However, owing to the uncertainties in the factors which contribute to lectin-induced cell agglutination (10), the molecular basis for these observations remain to be determined. In this study, we have used microspheres (11-14) coupled to either Con A or wheat germ agglutinin (WGA) as visual markers to study by scanning electron microscopy the topographical distribution of lectin receptors on D. discoideum cells fixed at different stages of development. We also describe the effect of labeling on the distribution of lectin receptors and on the morphology of the cell surface.

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

Parasitology ◽

10.1017/s0031182099004965 ◽

1999 ◽

Vol 119 (5) ◽

pp. 491-501 ◽

Cited By ~ 13

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.

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