The distribution of Concanavalin A receptor sites on the membrane of chromaffin granules

1975 ◽  
Vol 19 (1) ◽  
pp. 33-54
Author(s):  
P.A. Eagles ◽  
L.N. Johnson ◽  
C. Van Horn
Keyword(s):  
Concanavalin A ◽  
Plasma Membranes ◽  
Asymmetric Distribution ◽  
Chromaffin Granules ◽  
Intracellular Membrane ◽  
Chromaffin Granule ◽  
Binding Studies ◽  
Cell Cytoplasm ◽  
Con A ◽  
Receptor Sites

The distribution of concanavalin A (con A) receptor sites on the membranes of chromaffin granules has been investigated by binding studies using 125I-labelled con A and by electron-microscope studies using ferritin-labelled con A. In both experiments con A was observed to bind to chromaffin granule membranes but not to intact granules. The ferritin-con A particles bind to only one of the two possible surfaces of the chromaffin granule membranes. These results are in agreement with previous observations concerning the asymmetric distribution of saccharide residues on the surfaces of a number of different plasma membranes. They suggest that for the intracellular membrane of the chromaffin granule the saccharide sites, like those in plasma membranes, are not exposed to the cell cytoplasm. Further work is necessary to establish whether these sites are on the inner surface of the membrane or whether they are unmasked during the conversion of granules to membrane ghosts.

scholarly journals Cell surface-mediated activation of progelatinase A: demonstration of the involvement of the C-terminal domain of progelatinase A in cell surface binding and activation of progelatinase A by primary fibroblasts

1994 ◽  
Vol 304 (1) ◽  
pp. 263-269 ◽  
Author(s):  
R V Ward ◽  
S J Atkinson ◽  
J J Reynolds ◽  
G Murphy
Keyword(s):  
Cell Surface ◽  
Concanavalin A ◽  
Plasma Membranes ◽  
Scatchard Analysis ◽  
Gelatinase A ◽  
Surface Binding ◽  
Binding Studies ◽  
Cell Surface Binding ◽  
Terminal Domain ◽  
Primary Fibroblasts

We report that the isolated C-terminal domain of progelatinase A is inhibitory to the activation of this proenzyme by primary skin fibroblast plasma membranes but is unable to inhibit organomercurial-induced self-cleavage and activation. Ligand binding studies demonstrate that fibroblasts stimulated with concanavalin A to activate progelatinase A have a significantly enhanced level of cell surface-associated progelatinase A. Tissue inhibitor of metalloproteinases-2 (TIMP-2), an effective inhibitor of membrane-mediated progelatinase A activation, is able to abolish the enhanced level of cell surface-associated progelatinase A that occurs following stimulation. TIMP-1, a poor inhibitor of membrane activation, is unable to inhibit the cell surface binding of progelatinase A. The enhancement in the binding of 125I-progelatinase A to fibroblasts following concanavalin A stimulation can be blocked by the inclusion of excess C-terminal gelatinase A but not by a truncated form of gelatinase A lacking the C-terminal domain. Scatchard analysis of the binding of 125I-progelatinase A to concanavalin A-stimulated fibroblasts has identified 950,000 gelatinase binding sites per cell with a Kd of 1.3 x 10(-8) M. Analysis of non-stimulated fibroblasts has identified 500,000 sites per cell with a Kd of 2.6 x 10(-8) M. We propose that membrane-mediated activation of progelatinase A involves binding of the proenzyme through its C-terminal domain to the cell surface and that TIMP-2 can inhibit activation by interaction with progelatinase A through the C-terminal domain, thus preventing binding of the proenzyme.

Adhesion of Phytophthora palmivora zoospores: detection and ultrastructural visualization of concanavalin A-receptor sites appearing during encystment

1975 ◽  
Vol 19 (1) ◽  
pp. 11-20
Author(s):  
V.O. Sing ◽  
S. Bartnicki-Garcia
Keyword(s):  
Electron Microscopy ◽  
Cell Surface ◽  
Concanavalin A ◽  
Solid Surfaces ◽  
Trypsin Digestion ◽  
Phytophthora Palmivora ◽  
Con A ◽  
Binding Material ◽  
Ultraviolet Microscopy ◽  
Receptor Sites

The binding of concanavalin A (Con A) to the cell surface of zoospores and cysts of Phytophthora palmivora was studied by radiometry (125I-Con A), ultraviolet microscopy (fluorescein-Con A) and electron microscopy peroxidase-diaminobenzidine technique). Zoospores were found to secrete during the early stages of encystment a Con A-binding material susceptible to trypsin digestion. This glycoprotein is contained in the so-called peripheral vesicles and is probably responsible for the adhesion of the encysting zoospores to solid surfaces.

Electron microscopic localization of concanavalin A receptor sites in pollen surface material after fixation with glutaraldehyde-cetylpyridinium chloride.

1984 ◽  
Vol 32 (8) ◽  
pp. 869-871 ◽  
Author(s):  
M Grote ◽  
H G Fromme
Keyword(s):  
Concanavalin A ◽  
Cetylpyridinium Chloride ◽  
Pollen Grains ◽  
Dense Material ◽  
Con A ◽  
Electron Microscopic ◽  
Electron Dense Material ◽  
Receptor Sites ◽  
Pollen Surface ◽  
Ultrathin Sections

Pollen from birch trees (Betula pendula) was fixed in glutaraldehyde containing 0.5% cetylpyridinium chloride (CPC), incubated with concanavalin A (Con A)-ferritin, postfixed in osmium, dehydrated, and embedded in Epon. On ultrathin sections, ferritin particles were observed closely associated with the electron-dense material precipitated by CPC on the surface of the pollen grains. Controls for CPC, which were fixed in glutaraldehyde alone, showed no electron-dense material on the surface. In controls for Con A, which were incubated in Con A-ferritin in the presence of the inhibitory sugar (alpha-methyl-D-mannopyranoside), no ferritin particles were observed. The above-described procedure thus allows the localization of sugar residues in highly soluble pollen wall glycoproteins.

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 Ultrastructural localization of concanavalin A and wheat germ agglutinin binding sites in adult and embryonic mouse myocardium.

1982 ◽  
Vol 30 (3) ◽  
pp. 193-200 ◽  
Author(s):  
D Gros ◽  
B Bruce ◽  
C E Challice ◽  
J Schrevel
Keyword(s):  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ultrastructural Localization ◽  
Embryonic Cells ◽  
Con A ◽  
Embryonic Mouse ◽  
Transverse Tubular System ◽  
Receptor Sites ◽  
Ultrathin Sections

The lectins, concanavalin A (Con A) and wheat germ agglutinin (WGA), have been used to localize with precision glycosyl residues in adult and embryonic mouse myocardium. They were detected by means of an affinity method using peroxidase and chitobiosylperoxidase, respectively, which then were revealed with 3,3'-diaminobenzidine and H2O2. Exhaustive controls have shown that the binding of Con A and WGA is reversible when experiments are performed with adult specimens (tissue blocks or ultrathin sections of glycol methacrylate-embedded material) or with isolated embryonic cells. Experiments carried out with tissue blocks from embryonic hearts have shown peroxidase binding. This finding is discussed on the basis of the presence of the endogenous lectin-like components in embryonic hearts. Results show that the surface of adult and embryonic myocardial cells specifically bind both Con A and WGA, thus indicating the presence of glycosyl residues similar to alpha-methyl-D-mannoside and N-acetyl-D-glucosamine. In adult heart the transverse tubular system was also labeled. The absence of Con A and WGA receptor sites in the gap junction regions was demonstrated by means of an electron microscope postembedding staining method.

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.

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.

scholarly journals Membrane differentiation of developing hemic cells of the bone marrow demonstrated by changes in concanavalin A surface labeling.

1979 ◽  
Vol 27 (11) ◽  
pp. 1413-1423 ◽  
Author(s):  
G A Ackerman ◽  
W H Freeman
Keyword(s):  
Bone Marrow ◽  
Concanavalin A ◽  
Bone Marrow Cells ◽  
Ultrastructural Localization ◽  
Spatial Arrangement ◽  
Membrane Glycoproteins ◽  
Con A ◽  
Monocytic Cell ◽  
Receptor Sites ◽  
Cell Series

The concanavalin A-gold labeled horseradish peroxidase (Con A-HRP-G) method has been employed in the ultrastructural localization of Con A surface receptor sites on glutaraldehyde-fixed normal human and guinea pig bone marrow cells. The number of gold particles per micron of cell surface was counted and data subjected to statistical analysis. All cells of the bone marrow exhibited Con A binding; however, the extent of surface labeling was dependent both on cell type and stage of differentiation. Distinctive modifications in mean surface density correlated with specific periods during the maturation of the erythrocytic, neutrophilic, eosinophilic and monocytic cell series. In several instances, the differentiative changes in surface Con A labeling proved to be species dependent. These observations are discussed in relationship to methodology and to potential changes in number and/or spatial arrangement of Con A receptor sites, primarily attributable to mannosyl and/or glucosyl residues associated with membrane glycoproteins and/or glycolipids of developing neutrophilic and erythrocytic cells.

scholarly journals 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.

1985 ◽  
Vol 33 (5) ◽  
pp. 384-388 ◽  
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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