Fractionation of large glycopeptides of human teratocarcinoma-derived cells by concanavalin A – Sepharose chromatography

1980 ◽  
Vol 58 (4) ◽  
pp. 281-286 ◽  
Author(s):  
Maija-Liisa Rasilo
Keyword(s):  
Concanavalin A ◽  
Gel Filtration ◽  
Neuraminic Acid ◽  
Void Volume ◽  
Con A ◽  
The Third ◽  
Large Size

Human teratocarcinoma derived cells, line PA 1, were labeled with radioactive monosaccharides and subsequently digested with pronase. Large sized glycopeptides (fraction A) were isolated by gel filtration on Bio-Gel P-10. Their chromatography on concanavalin A – Sepharose gave three subfractions, two of which were eluted with a sugar-free buffer and the third with 10 mM α-methyl mannoside. The first subfraction (fraction A – Con A Ia) incorporated label from [3H]galactose and [3H]glucosamine and contained the largest components of fraction A. The second and the third subfractions (fractions A – Con A Ib and A – Con A II) were glycopeptides which incorporated label from tritiated fucose, mannose, galactose, and glucosamine. Even these molecules were of large size eluting partially at the void volume from Bio-Gel P-60. The glycopeptides of fraction A – Con A Ib contained mannose, fucose, galactose, N-acetylglucosamine, and N-acetylgalactosamine. Fucose and galactose residues occupied ultimate or penultimate positions at the nonreducing termini of the oligosaccharides. N-Acetyl-neuraminic acid, too, was present in the glycopeptides of fraction A.

Partial structural analysis of concanavalin A binding glycopeptides of large size from human teratocarcinoma-derived cells: cleavage by hydrazinolysis and alkaline borohydride

1982 ◽  
Vol 60 (7) ◽  
pp. 749-756 ◽  
Author(s):  
Maija-Liisa Rasilo ◽  
Ossi Renkonen
Keyword(s):  
Structural Analysis ◽  
Concanavalin A ◽  
Relative Mass ◽  
Human Origin ◽  
Con A ◽  
Large Size ◽  
Teratocarcinoma Cells ◽  
The Individual ◽  
Mouse Teratocarcinoma

Pronase digests of cultured teratocarcinoma-derived cells (PA 1) of human origin have been previously shown to contain large-sized glycopeptides (relative mass (Mr) > 7400), of which 15–23% are retained by columns of concanavalin A (Con A) – Sepharose and can be eluted with 10 mM methyl α-D-mannopyranoside. The present data show that this fraction (A – Con A II) contains a family of glycopeptides that are degradable with anhydrous hydrazine as well as with 0.05 M NaOH – 1 M NaBH4. The cleavage products representing individual oligosaccharide chains, presumably as oligosaccharides and glycopeptides, consisted mostly of medium- (Mr 1400–6000) and small-sized (Mr < 1400) molecules. This implies that glycopeptides bearing several oligosaccharide chains were present in A – Con A II. Most of the individual oligosaccharide chains were not bound to Con A – Sepharose, but some were retained by the lectin column in the same way as the original glycopeptides. Some of the oligosaccharides were degraded partially with endo-β-galactosidase from Escherichia freundii suggesting the presence of GalβGlcNAcβ repeats. The present findings show that A – Con A II may be different from the "embryonic" glycopeptides of mouse teratocarcinoma cells that are reportedly not cleaved by mild alkaline borohydride treatment. Instead, A – Con A II is reminiscent of the T-1 glycopeptide of glycophorin.

scholarly journals THE EFFECT OF TOASTING, DRY UREA TREATMENT AND SPROUTING ON SOME THERMOSTABLE TOXIC FACTORS IN THE JACKBEAN SEED

2021 ◽  
Vol 25 (1) ◽  
pp. 36-39
Author(s):  
B. O. ESOUN ◽  
A. B. I. UDEDIBIE ◽  
C. R. CARLINI
Keyword(s):  
Concanavalin A ◽  
Crude Protein ◽  
Appreciable Effect ◽  
Con A ◽  
Feed Ingredient ◽  
Urea Treatment ◽  
The Third ◽  
Toxicological Studies ◽  
Ruminant Animals ◽  
Toxic Factors

Raw unprocessed jackbean seed contains 26 – 32% crude protein and also toxic elements most of which are thermostable, which limit its use as feed ingredient for livestock especially non-ruminant animals. Raw jackbean seeds were divided into three batches. One batch was ground and toasted, the second, batch was ground raw and mixed with 2% of its weight of dry area and allowed to stand for 11days. The third batch was sprouted for four days and later ground into meal. Toxicological studies on the batches of the jackbean meals were conducted for concanavalin A (Con A) and canatoxin in jackbean seed, while sprouting was effective in detoxifying concanavalin A (Con A) and canatoxin but not very effective in detoxifying the urease activity in jackbean seed. Toasting alone did not have appreciable effect on these toxic factors

MONOCLONAL ANTIBODY WITH PREFERENTIAL AFFINITY FOR LARGE MULTIMERS OF VON WILLEBRAND FACTOR

1987 ◽  
Author(s):  
Tomiko Ryu ◽  
Akiko Nakayama ◽  
Atsushi Oguchi ◽  
Tadatoshi Kinoshita ◽  
Mutsuyoshi Kazama ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Von Willebrand Factor ◽  
Gel Filtration ◽  
Void Volume ◽  
Size Fractions ◽  
Large Size ◽  
Function Relationship ◽  
Von Willebrand ◽  
Relationship Of ◽  
Willebrand Factor

von Willebrand factor (vWF) consists of a series of multimers of 270,000 mol. wt. subunits. Ristocetin cofactor activity (vWF:RCo) and capacity of binding to platelets of vWF are associated with large multimers, and Type IIA von Willebrand's disease (vWD) is characterized by lack of the large multimers. The significance of multimeric structure in relation to vWF function remains unclear. We obtained a monoclonal antibody (MAb) to human vWF which inhibited ristocetin-induced platelet aggregation. This antibody proved to bind preferentially to larger multimers by the finding that the MAb-conjugated Sepharose adsorbed large multimers of vWF from cryoprecipitates, leaving small multimers unadsorbed. vWF:Ag levels of noraml subjects determined by ELISA using the MAb correlated well with those by ELISA using polyclonal antibody (PAb) to vWF. When the plasma from patients with Type IIA vWD and platelet type vWD were examined, the values obtained by the MAb ELISA had a good correlation with vWF:RCo, but were lower than the values obtained by the PAb ELISA. In gel filtration of factor VIII concentrate, vWF:Ag detected by the MAb and vWF:RCo were present in the void volume and large size fractions, whereas vWF:Ag detectable with PAb appeared broadly from the void volume to smaller size fractions. The MAb inhibited ristocetin-dependent binding of vWF to platelets, but did not affect ADP-induced binding of vWF to platelets. These findings suggest that the large multimers have a function-associated specific structure which is absent in the small multimers, and the MAb will be useful for the investigation of multimer-function relationship of vWF.

A structural basis for four distinct elution profiles on concanavalin A – Sepharose affinity chromatography of glycopeptides

1979 ◽  
Vol 57 (1) ◽  
pp. 83-96 ◽  
Author(s):  
Saroja Narasimhan ◽  
James R. Wilson ◽  
Eva Martin ◽  
Harry Schachter
Keyword(s):  
Affinity Chromatography ◽  
Weak Interaction ◽  
Concanavalin A ◽  
Tight Binding ◽  
Structural Basis ◽  
Con A ◽  
Mannose Residue ◽  
Terminal Residue ◽  
The Third ◽  
Profile Type

Twelve 14C-acetylated glycopeptides have been subjected to affinity chromatography on concanavalin A (Con A) – Sepharose at pH 7.5. The elution profiles could be classified into four distinct patterns. The first pattern showed no retardation of glycopeptide on the column and was elicited with a glycopeptide having three peripheral oligosaccharide chains:[Formula: see text]Such glycopeptides have only a single mannose residue capable of interacting with Con A – Sepharose; an interacting mannose residue is either an α-linked nonreducing terminal residue or an α-linked 2-O-substituted residue. The second type of profile showed a retarded elution of glycopeptide with buffer lacking methyl α-D-glucopyranoside (indicative of weak interaction with the column) and was given by glycopeptides with the structures:[Formula: see text]where R1 is either H or a sialyl residue. The third profile type showed tight binding of glycopeptide to Con A – Sepharose and elution as a sharp peak with 0.1 M methyl α-D-glucopyranoside; glycopeptides giving this pattern had the structures:[Formula: see text]where R2 is either H, GlcNAc, Gal-β1,4-GlcNAc, or sialyl-Gal-β1,4-GlcNAc. These glycopeptides all have two interacting mannose residues, the minimum required for binding to the column; one of these mannose residues must, however, be a terminal residue to obtain tight binding and sharp elution. The fourth profile type showed tight binding of glycopeptide to the column but elution with 0.1 M methyl α-D-glucopyranoside resulted in a broad peak indicating very tight binding; glycopeptides showing this behaviour had the structures:[Formula: see text]where R3 is either GlcNAc, Gal-β1,4-GlcNAc, or siaryl-Gal-β1,4-GlcNAc. Therefore it can be concluded that although a minimum of two interacting mannose residues is required for binding to Con A – Sepharose, the residues linked to these mannoses can either strengthen or weaken binding to the column.

Effects of concanavalin A on developing ganglion cells in the retina of chick embryos

Development ◽  
1981 ◽  
Vol 65 (1) ◽  
pp. 27-39
Author(s):  
K. Meller
Keyword(s):  
Cell Death ◽  
Concanavalin A ◽  
Ganglion Cells ◽  
Positional Information ◽  
Chick Embryos ◽  
Con A ◽  
The Third ◽  
Receptor Sites ◽  
Retinal Neurogenesis ◽  
Natural Cell

The administration of concanavalin A (Con A) (50–200 µg/egg) to chick embryos between the third and the seventh day of incubation has the following effects on the retina: (1) Con A causes the degeneration of a large number of ganglion cells and consequently the layer that should be formed by these cells is not present or is constituted only by a small number of ganglion cells. (2) The lectin seems to be effective only when it is administered during the postmitotic phase of the ganglion cells. (3) The degenerated cells are phagocytosed by the Müller cells in a manner similar to that occurring during the natural cell death in normal retinal development. (4) The differentiation of other retinal elements (photoreceptors, bipolar, amacrine and Müller cells) is not affected by the lectin administration. (5) The administration of Con A in later stages of development, even at ten times higher dosages (2000 µg/egg), fails to affect retinal neurogenesis. It is suggested that Con A binding to receptor sites of the cell membrane affects the distribution or mobility of surface components producing an alteration in the mechanism by which the developing cells regulate positional information during retinal neurogenesis.

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.

scholarly journals Jaws of a large belemnite and an ammonite from the Aalenian (Middle Jurassic) of Switzerland

2020 ◽  
Vol 139 (1) ◽  
Author(s):  
Christian Klug ◽  
Walter Etter ◽  
René Hoffmann ◽  
Dirk Fuchs ◽  
Kenneth De Baets
Keyword(s):  
Middle Jurassic ◽  
Opalinus Clay ◽  
Northern Switzerland ◽  
The Third ◽  
Large Size ◽  
Upper Jaw ◽  
Rule Out

AbstractAlthough belemnite rostra can be quite abundant in Jurassic and Cretaceous strata, the record of belemnite jaws was limited to a few specimens from Germany and Russia. Here, we describe and figure three cephalopod jaws from the Middle Jurassic Opalinus Clay of northern Switzerland. Although flattened, the carbonaceous fossils display enough morphological information to rule out an ammonoid, nautiloid or octobrachian origin of the two larger jaws. Their similarities to belemnite jaws from Germany and Russia conforms with our interpretation of these specimens as belemnite jaws. Based on their rather large size, we tentatively assign these two jaws to the megateuthidid Acrocoelites conoideus. The third jaw is a rather small upper jaw of an ammonoid. Since Leioceras opalinum is by far the most common ammonite in this unit in northern Switzerland, we tentatively suggest that the upper jaw belongs to this species.

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.

Acid proteases from species of Mucor. III. Interaction with concanavalin A and concanavalin A Sepharose

1976 ◽  
Vol 54 (2) ◽  
pp. 120-129 ◽  
Author(s):  
W. S. Rickert ◽  
P. A. McBride-Warren
Keyword(s):  
Molecular Weight ◽  
Concanavalin A ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Affinity Column ◽  
Mucor Miehei ◽  
Highly Active ◽  
Elution Buffer ◽  
Turbidimetric Assay ◽  
Ph Range

The reaction of Mucor miehei protease with concanavalin A was followed by a turbidimetric assay in the pH range 5–8. At pH 4.0, no turbidity developed but binding of the enzyme to concanavalin A could be demonstrated by gel filtration. Two fractions of apparent molecular weight 65 000 and 52 000 were isolated, the 65 000 molecular weight species apparently representing a protomer of concanavalin A (24 000) bound to the enzyme. An analysis of the circular dichroism spectrum of this complex suggested that protomer binding results in a conformational change in the enzyme which is associated with a 30% increase in proteolytic activity.At pH 6.0, the enzyme was strongly bound to columns of concanavalin A Sepharose but could be removed by including α-methyl D-glucoside and NaCl in the elution buffer. Some column degradation occurred at room temperature but was not detectable at 4 °C where rapid elution of the enzyme resulted in a greater than 90% yield of highly active protein. Periodate-oxidized Mucor miehei protease and Mucor rennin did not react with concanavalin A and were not bound to the affinity column.

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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