Leguminous Seed Glycoproteins That Interact With Concanavalin A

1977 ◽  
Vol 4 (1) ◽  
pp. 25 ◽  
Author(s):  
PA Gleeson ◽  
MA Jermyn
Keyword(s):  
Concanavalin A ◽  
Coffea Arabica ◽  
Sodium Dodecyl ◽  
Con A ◽  
Precipitation Line ◽  
Leguminous Seed ◽  
Protein Staining ◽  
Gel Diffusion ◽  
Haemagglutination Activity ◽  
Seed Extracts

Material that interacts with concanavalin A has been purified from seed extracts of 18 species (16 legumes, Coffea arabica and Lolium perenne) by Con A-Sepharose chromatography. When examined by sodium dodecyl sulphate-polyacrylamide disc gel electrophoresis, many of the preparations showed a number of protein-staining components. All the preparations gave a single precipitation line against Cunalialia ensiformis crude extract on gel diffusion plates. Some of the con A-interacting preparations were shown to possess haemagglutination activity. The Arachis hypogaea (peanut) preparation contains one major glycoprotein of molecular weight 69 000, which was further purified and analysed. The purified glycoprotein contains 12% carbohydrate, galactose being the major sugar with minor amounts of mannose and xylose present.

Precipitation Reactions Between Components of Plant Tissue Extracts

1975 ◽  
Vol 2 (4) ◽  
pp. 533 ◽  
Author(s):  
MA Jermyn
Keyword(s):  
Concanavalin A ◽  
Carbohydrate Binding ◽  
Exact Nature ◽  
Plant Seeds ◽  
Tissue Extracts ◽  
General Inhibitor ◽  
Precipitation Reactions ◽  
Leguminous Seeds ◽  
Gel Diffusion ◽  
Seed Extracts

Precipitation lines are observed on gel-diffusion plates between many pairs of seed extracts. Interactions involving Canavalia ensiformis depend upon concanavalin A, and a number of extracts from other plant seeds contain a precipitant that mimics that lectin. Methyl a-D-mannopyranoside is a general inhibitor of the precipitation phenomenon and some form of protein-carbohydrate binding seems to be involved in all precipitations, although the exact nature of the macromolecules taking part is obscure. For the leguminous seeds Phaseolus vulgauis, Vicia faba and Cajanus cajan, precipitation reactions occur between extracts of cotyledons and extracts of tissues of the parent plants, even of the testa of the seeds. The nature of these reactions appears to be the same as those of the interspecies ones. Both types of reaction may be examples of ways in which plant cells recognize self from non-self. The material in P. vulgaris that reacts with concanavalin A is a group of globulin-like glycoproteins (5 % carbohydrate), heterogeneous in both charge and molecular weight but similar in overall amino acid analysis.

Identification of liver plasma membrane glycoproteins which bind to 125I-Labelled concanavalin A following electrophoresis in sodium dodecyl sulfate

1976 ◽  
Vol 54 (5) ◽  
pp. 477-480 ◽  
Author(s):  
James W. Gurd ◽  
W. Howard Evans
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Concanavalin A ◽  
Gel Electrophoresis ◽  
Plasma Membranes ◽  
Sodium Dodecyl ◽  
Membrane Composition ◽  
Membrane Glycoproteins ◽  
Con A ◽  
General Application ◽  
Dodecyl Sulfate

Following electrophoresis of ovalbumin in sodium dodecyl sulfate (SDS) this glycoprotein bound 125I-labelled concanavalin A (Con A). The reaction was specific and proportional to the amount of glycoprotein present on the gel. This technique was used to study the Con-A-binding glycoproteins of liver cell surfaces. Mouse liver plasma membranes were purified and subfractionated to yield two fractions corresponding to the bile canalicular surface and the surface between adjacent hepatocytes (Evans, W. H. (1970) Biochem. J. 116, 833–842). Both fractions bound 125I-labelled Con A, the former binding two to three times more lectin than the latter. Following SDS gel electrophoresis individual membrane glycoproteins reacted with 125I-labelled Con A. Both membrane subfractions yielded qualitatively similar Con A binding profiles, seven binding proteins being present in each. The results are consistent with a generally uniform distribution of glycoproteins over the hepatocyte surface. The reaction of lectins with glycoproteins following SDS gel electrophoresis should find general application in the study of membrane composition.

scholarly journals A study of maturation events in jackbeans (Canavalia ensiformis)

1984 ◽  
Vol 222 (1) ◽  
pp. 265-268 ◽  
Author(s):  
S E Marcus ◽  
J Burgess ◽  
P R Maycox ◽  
D J Bowles
Keyword(s):  
Concanavalin A ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Molecular Forms ◽  
Con A ◽  
Canavalia Ensiformis ◽  
Immature Seeds

Maturation events have been studied in developing jackbean (Canavalia ensiformis) cotyledons by using a combination of analysis by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, overlays with 125I-concanavalin A (Con A) and the use of anti-(Con A) after Western transfer. The number of polypeptides recognized by 125I-Con A varies during maturation, until at maturity only one remains. Several molecular forms of the lectin occur during development; one, corresponding to Mr 33 000 and found only in immature seeds, interacts with 125I-Con A, suggesting that it is glycosylated.

scholarly journals Identification of concanavalin A receptors and galactose-binding proteins in purified plasma membranes of Dictyostelium discoideum.

1977 ◽  
Vol 74 (1) ◽  
pp. 264-273 ◽  
Author(s):  
C M West ◽  
D McMahon
Keyword(s):  
Dictyostelium Discoideum ◽  
Concanavalin A ◽  
Plasma Membranes ◽  
Sodium Dodecyl ◽  
Fluorescein Isothiocyanate ◽  
Binding Activity ◽  
Cellular Slime Mold ◽  
Con A ◽  
Stage Of Development ◽  
Cellular Slime

Two techniques have been modified to provide simple means for the identification of molecules which bind concanavalin A (Con A). Crossed immunoelectrophoresis was altered by replacing antibody with Con A, and receptors were identified by the precipitin arcs which they produced. Con A, tagged with fluorescein isothiocyanate, was also diffused into prefixed sodium dodecyl sulfate (SDS)-polyacrylamide gels, and additional receptors identified by fluorescence. More than 35 molecules in the plasma membranes of the cellular slime mold Dictyostelium discoideum which bind Con A were identified with these techniques. At least 12 of these diminish and 12 increase in importance as receptors during differentiation of the cells from the vegetative to the preculmination stage of development. In the course of these experiments, it was possible to confirm the presence of the galactose-binding protein discoidin, in the plasma membrane, by electrophoresing membrane proteins into an agarose gel. This lectin regains its sugar-binding activity after denaturation and electrophoresis in SDS.

Changes in concanavalin A-reactive proteins in inflammatory disorders.

1989 ◽  
Vol 35 (11) ◽  
pp. 2207-2211 ◽  
Author(s):  
B Silvestrini ◽  
A Guglielmotti ◽  
L Saso ◽  
C Y Cheng
Keyword(s):  
Autoimmune Diseases ◽  
Concanavalin A ◽  
Lupus Erythematosus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Con A ◽  
Serum Samples ◽  
Inflammatory Disorders ◽  
Reducing Conditions ◽  
Human Sera

Abstract Quantitative changes of concanavalin A (Con A)-reactive proteins in serum samples obtained from rats with induced inflammation and from patients with inflammatory and autoimmune diseases were examined by use of lectin blots. Treatment of rats with a single dose of fermented yeast to induce inflammation caused an extensive increase in Con A-reactivity. These changes were time dependent and were similar in both sexes of the animals. When we examined serum samples obtained from patients with various inflammatory disorders for their Con A-reactive proteins as compared with normal donors, we noted that the Con A-reactivity increased in patients with rheumatoid arthritis and systemic lupus erythematosus. Among all the glycoproteins examined by lectin blots with use of Con A, a set of five proteins was selected for detailed analysis by densitometric scanning. These included alpha 2-macroglobulin, P-150, P-95, P-40, and P-35, of Mr 180,000, 150,000, 95,000, 40,000, and 35,000, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Densitometric scanning analysis of the lectin blots revealed that the Con A-reactivity of these proteins increased during inflammation. Because alpha 2-macroglobulin is not an acute-phase protein in humans, an increase in Con A staining of this protein suggested that altered glycation is associated with autoimmune diseases. Thus, study of changes in Con A-reactive proteins in human sera may facilitate our understanding of the etiology and pathophysiology of autoimmune diseases.

Concanavalin A-induced Aggregation of Human Blood Platelets

1975 ◽  
Vol 33 (02) ◽  
pp. 354-360 ◽  
Author(s):  
Heinrich Patscheke ◽  
Reinhard Brossmer
Keyword(s):  
Concanavalin A ◽  
Binding Capacity ◽  
Specific Binding ◽  
Blood Platelets ◽  
Residual Effect ◽  
Independent Effect ◽  
Con A ◽  
Experimental Conditions ◽  
Main Effect ◽  
Induced Aggregation

SummaryConcanavalin A (CON A) causes platelets to aggregate. A Ca++-independent effect of CON A could be separated from a main effect which depends on Ca++. The main effect probably is a consequence of the CON A-induced platelet release reaction and therefore is platelet-specific. The weak residual effect observed in the presence of Na2EDTA may be due to a similar mechanism as has been demonstrated for CON A-induced aggregations of several other normal and malignant transformed animal cells.Na2EDTA did not inhibit the carbohydrate-specific binding capacity of CON A. Therefore, Na2EDTA appears not to demineralize the CON A molecules under these experimental conditions.α-methyl-D-glucoside inhibits the Ca++-independent as well as the Ca++-dependent effect of CON A.Pretreatment by neuraminidase stimulated the platelet aggregation induced by CON A. It is possible that removal of terminal sialic acid residues makes additional receptors accessible for the binding of CON A.

Immunological Determined Plasminogen Groups And Its Possible Biological Relevance

1981 ◽  
Author(s):  
V Sachs ◽  
R Dörner ◽  
E Szirmai
Keyword(s):  
Human Plasma ◽  
Type I ◽  
Type Ii ◽  
Precipitation Line ◽  
Gene Frequencies ◽  
Type Iii ◽  
Different Types ◽  
Human Plasminogen ◽  
Gel Diffusion ◽  
Good Agreement

Anti human plasminogen sera of the rabbit precipitate human plasma in the agar gel diffusion test by means of intra-basin absorption with plasminogenfree human plasma with three different types: type I is represented by one strong precipitation line, type II by two lines, a big one and a small one, and type III by three slight but distinct lines. The following frequencies of the different types have been observed in a sample of 516 human plasmas: type I 65%, type II 33% and type III 2%. Suppose the types are phenotypical groups of a diallelic system where the types I and III represent the homozygous genotypes and the type II the heterozygous the estimated gene frequencies are in good agreement with the expected values. There is also a good agreement of the distribution of plasminogen groups determined by electrofocussing from RAUM et al. and HOBART. The plasminogen groups possibly may have also a biological meaning because the plasmas of type III always have a lesser fibrinolytic activity than the plasmas of the other types.

scholarly journals Visualization of intracellular concanavalin A binding sites in retinal photoreceptors.

1978 ◽  
Vol 26 (10) ◽  
pp. 822-828 ◽  
Author(s):  
I Nir
Keyword(s):  
Concanavalin A ◽  
Binding Sites ◽  
Photoreceptor Cells ◽  
Con A ◽  
Thin Sections ◽  
Glycol Methacrylate ◽  
Outer Segments ◽  
Retinal Photoreceptors ◽  
Carbohydrate Components ◽  
The Relationship

Localization of carbohydrate components in retinal photoreceptor cells and membranes was studied. Frog and rat retinas were fixed with glutaraldehyde and embedded in glycol methacrylate or in a mixture of glycol methacrylate, glutaraldehyde and urea. Thin sections were incubated with ferritin-labeled concanavalin A (F-Con A) and stained with osmium vapors. Intensive binding was observed in both rod and cone outer segments. In the rod inner segment, differential binding of F-Con A was demonstrated. While numerous ferritin granules were observed in the myoid zone, only a few were seen in the ellipsoid zone, except for a local accumulation along the plasma membrane. In the rod outer segment, Con A binding sites were closely associated with the disk membranes. Ferritin granules were observed on both sides of the membranes. The relationship between the localization of Con A binding sites and the orientation of visual pigment molecules within the rod outer segments disk membranes was discussed.

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

1992 ◽  
Vol 38 (8) ◽  
pp. 1418-1424 ◽  
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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