Endogenous cell surface lectin in Dictyostelium: quantitation, elution by sugar, and elicitation by divalent immunoglobulin.

Springer, R. W.; Haywood, Patricia L.; Barondes, Samuel H.

doi:10.1083/jcb.87.3.682

Cited by 40 publications

(27 citation statements)

References 14 publications

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“…While the number of cell surface discoidins has not been definitively established [ 10,13,26], the available binding data suggest a lower affinity of antidiscoidin antibodies for their antigens in intact cells, as compared to antibodies directed to other cell adhesion molecules [ 131.…”

Section: Discussionmentioning

confidence: 99%

The role of membrane lectins in dictyostelium discoideum aggregation as ascertained by specific univalent antibodies against discoidin I

Cano

Pestaña

1984

J of Cellular Biochemistry

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Antibodies against pure discoidin I have been used as a tool to ascertain the role of this lectin in aggregation of Dictyostelium discoideum. Discoidin I is widely expressed over the cell surface of aggregation-competent AX-2 cells, as ascertained by indirect immunofluorescence with specific (antidiscoidin I) antibodies. Univalent antidiscoidin I antibodies (Fab fragments) inhibit the aggregation-specific intercellular adhesion of D discoideum AX-2 cells in an in vitro assay. This inhibition depends on antibody concentration and cell density; a 50% inhibition of cell aggregation was obtained at antidiscoidin I Fab concentration of 4.5 mg/ml and 1 X 10(6) cells/ml. Aggregation and morphogenesis on solid support is also effectively inhibited when AX-2 cells are starved in the presence of antidiscoidin I Fab fragments. The inhibition of morphogenesis is also dose dependent and more effective than in the in vitro assay. No inhibition of aggregation either in the in vitro assay or on morphogenesis on solid support was observed with preimmune Fab fragments at any of the concentrations tested (up to 9.6 mg/ml).

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Section: Discussionmentioning

confidence: 99%

The role of membrane lectins in dictyostelium discoideum aggregation as ascertained by specific univalent antibodies against discoidin I

Cano

Pestaña

1984

J of Cellular Biochemistry

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“…The existence of membrane and soluble forms of surface antigens has also been seen for the variant surface glycoproteins of trypanosomes where the membrane-bound form has an anchoring phospatidylinositol group (25), and the cellular slime mold-soluble lectin that binds to a galactose-containing receptor on the slime mold surface (26). The biochemical differences and the biologic significance of the membrane-bound and soluble forms of the amebic lectin are under investigation.…”

Section: Discussionmentioning

confidence: 99%

Isolation of the galactose-binding lectin that mediates the in vitro adherence of Entamoeba histolytica.

Petri¹,

Smith²,

Schlesinger³

et al. 1987

J. Clin. Invest.

274

193

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Entamoeba histolytica adheres to human colonic mucus, colonic epithelial cells, and other target cells via a galactose (Gal) or N-acetyl-D-galactosamine (GalNAc) inhibitable surface lectin. Blockade of this adherence lectin with Gal or GalNAc in vitro prevents amebic killing of target cells. We have identified and purified the adherence lectin by two methods: affinity columns derivatized with galactose monomers or galactose terminal glycoproteins, and affinity columns and immunoblots prepared with monoclonal antibodies that inhibit amebic adherence. By both methods the adherence lectin was identified as a 170-kD secreted and membrane-bound amebic protein. The surface location of the lectin was confirmed by indirect immunofluorescence. Purified lectin competitively inhibited amebic adherence to target cells by binding to receptors on the target Chinese hamster ovary cells in a Gal-inhibitable manner.

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“…Nevertheless, we have determined ( Table I) that approximately 3 x lo5 molecules of Protein A are bound to cells saturated with antidiscoidin (1 pl antiserum under these conditions). This value is comparable to the number of surface discoidin molecules obtained by competition radioimmunoassays with intact cells (2 x lo5) [14], but considerably higher than those obtained either by selective elution of discoidin from the cell surface (0.52 x lo5) or by direct binding of radioiodinated monovalent antidiscoidin Fab (0.37 x lo5) [15]. It should be noted that in a related species of slime mold, widely divergent estimates have been obtained using different immunochemical procedures to measure absolute quantities of a surface-localized lectin (purpurin) [15].…”

Section: Discussionmentioning

confidence: 65%

Immunochemical Analysis of Discoidins I and II at the Cell Surface in Wild Type and Aggregation‐Defective Mutants of Dictyostelium discoideum

Armant¹,

Berger²

1982

J of Cellular Biochemistry

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The endogenous lectins discoidins I and II are believed to be primary components of the morphogenetic cell cohesion system of D discoideum. We have developed two immunochemical methods to analyze the association of the discoidins with the cell surface. One method is a two-state specific antibody binding assay in which intact cells are incubated on ice with rabbit serum (either control serum or antidiscoidin I and II), washed, then incubated with 125I-Protein A. Specific antibody binding is defined as the difference between percent radioactivity bound with antidiscoidin versus control serum during the first stage. Substantial specific binding was observed with developed A3 cells but not with vegetative cells, and nearly all of the activity could be removed by preadsorption of the antiserum with discoidin-Sepharose. As a complementary method, quantitative immunoadsorption analysis was performed in which we tested the ability of intact cells to remove antibodies reactive with purified 125I-discoidin I or II. Developed cells, but not vegetative cells, were capable of adsorbing antibodies reactive with discoidin I as well as those reactive with discoidin II. This represents the first demonstration that both lectins are present on the surface of cohesive cells. These procedures, coupled with other methods to analyze soluble discoidin in cell extracts, were used to study discoidin expression in wild type cells and in two newly isolated aggregation-defective mutants. Strain EB-32 fails to aggregate and displays little or no discoidin in cell extracts or at the cell surface. On the other hand, strain EB-18 forms loose amorphous mounds, and expresses substantial quantities of the discoidins, both in cell extracts and at the cell surface. These mutants should prove valuable in studying the organization and regulation of discoidins I and II at the surface of aggregating cells.

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Endogenous cell surface lectin in Dictyostelium: quantitation, elution by sugar, and elicitation by divalent immunoglobulin.

Cited by 40 publications

References 14 publications

The role of membrane lectins in dictyostelium discoideum aggregation as ascertained by specific univalent antibodies against discoidin I

The role of membrane lectins in dictyostelium discoideum aggregation as ascertained by specific univalent antibodies against discoidin I

Isolation of the galactose-binding lectin that mediates the in vitro adherence of Entamoeba histolytica.

Immunochemical Analysis of Discoidins I and II at the Cell Surface in Wild Type and Aggregation‐Defective Mutants of Dictyostelium discoideum

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