Inhibition of intercellular adhesion in a cellular slime mould by univalent antibody against a cell-surface lectin

Rosen, Steven D.; Haywood, Patricia L.; Barondes, Samuel H.

doi:10.1038/263425a0

Cited by 39 publications

(4 citation statements)

References 13 publications

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“…Although considerable evidence has been presented that discoidin plays a role in cell-cell adhesion in D. discoideum (1,11), that role is not apparently reflected in the assay of cell-cell adhesion used in the present experiments . Except under unusual assay conditions, we (12) and others (13) have failed to inhibit cell-cell adhesion of another species of slime mold with univalent antibody fragments directed against its cell surface lectin . Whatever the significance of these results, they support the major point of this paper, the consistency of results with the present technique and with univalent antibody fragments made from the primary antibody .…”

Section: Resultsmentioning

confidence: 88%

Cell adhesion molecules: detection with univalent second antibody.

Springer¹,

Barondes²

1980

The Journal of Cell Biology

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Identification of cell surface molecules that play a role in cell-cell adhesion (here called cell adhesion molecules) has been achieved by demonstrating the inhibitory effect of univalent antibodies that bind these molecules in an in vitro assay of cell-cell adhesion . A more convenient reagent, intact (divalent) antibody, has been avoided because it might agglutinate the cells rather than blocking cell-cell adhesion . In this report, we show that intact rabbit immunoglobulin directed against certain cell surface molecules of Dictyostelium discoideum blocks cell-cell adhesion when the in vitro assay is performed in the presence of univalent goat anti-rabbit antibody . Under appropriate experimental conditions, the univalent second antibody blocks agglutination induced by the rabbit antibody without significantly interfering with its effect on cell-cell adhesion . This method promises to be useful for screening monoclonal antibodies raised against potential cell adhesion molecules because: (a) it allows for the screening of large numbers of antibody samples without preparation of univalent fragments; and (b) it requires much less antibody because of the greater affinity of divalent antibodies for antigens .

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Section: Resultsmentioning

confidence: 88%

Cell adhesion molecules: detection with univalent second antibody.

Springer¹,

Barondes²

1980

The Journal of Cell Biology

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“…These carbohydrate-binding proteins, the lectins, have very high specificity to their binding moieties. 99 Specific interactions between lectins and carbohydrates are essential in many biological processes, such as regulation of cell adhesion, 100,101 metastasis, 102 immune response, 103,104 and pathogen–host infections. 105 Carbohydrate microarrays composed of many distinct glycans immobilized on solid substrates provide ways to simultaneously detect multiple bindings of proteins, viruses, and cells to every glycan under identical conditions.…”

Section: Microarraysmentioning

confidence: 99%

Use of Microarrays as a High-Throughput Platform for Label-Free Biosensing

Sun

2015

SLAS Technology

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In recent years, various label-free biosensing technologies have been developed for studying the real-time kinetics of diverse biomolecular interactions. These biosensors partially take the place of fluorescence-based methods by providing a comparable sensitivity as well as retaining the conformational and functional integrality of biomolecules to be investigated. However, to completely eliminate the need of fluorescence, throughput is the next big consideration. Microarrays provide a high-throughput platform for screening tens of thousands of biomolecular interactions simultaneously, and many compatible fluorescent scanners have been commercially available. The combination of microarrays and label-free biosensors will be of great interest to researchers in related fields. Microarrays are fabricated by spotting, imprinting, or directly synthesizing biomolecules on solid supports such as glasses, silicon wafers, and other functionalized substrates, and they have been applied to detect DNAs, proteins, toxins, and so on in surface plasmon resonance (SPR) imaging systems and oblique-incidence reflectivity difference (OI-RD) microscopes. Current challenges include increasing sensitivity, reducing sampling time, improving surface chemistry, identifying captured molecules, and minimizing reagent consumption. Future research directions are to improve the instruments themselves, modify the microarray surface for more efficient analyte capture, and combine the systems with mass spectrometry and microfluidics.

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“…Since the 3H from the acetate is incorporated into carbohydrates and proteins, the equivalence of the two ratios suggests that the HAI, rather than being a modified food product, is synthesized de novo from the same pool of precursors that go into other slime mold proteins or carbohydrates. FIGURE 6 Cellulose acetate electrophoresis of labeled HAI . A pool (designated by the bar in Fig .…”

Section: Release Of Hai Into Mediummentioning

confidence: 99%

Identification and purification of an endogenous receptor for the lectin pallidin from Polysphondylium pallidum.

Drake¹,

Rosen²

1982

The Journal of Cell Biology

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We report the identification and purification of an endogenous carbohydratecontaining receptor of pallidin, the cell surface lectin implicated in mediating cell-cell adhesion in the cellular slime mold Polysphondylium pallidum . The receptor is identified in an aqueous extract of crude P. pallidurn membranes as a potent inhibitor of the hemagglutination activity of pallidin . The inhibitor is purified to apparent homogeneity by affinity precipitation with pallidin followed by fractionation of the solubilized precipitate on Sepharose 4B . The hemagglutination inhibitor (HAI) is metabolically radiolabeled, indicating that it is a biosynthetic product of the amoebae and not an ingested food substance. The HAI is released into the extracellular medium by living, differentiated amoebae. This release is markedly facilitated by the addition of D-galactose, a specific saccharide that binds to pallidin . Hence, the HAI appears to have an in situ association with pallidin at the cell surface. Exogenously added HAI promotes the agglutination of differentiated amoebae in a gyrated suspension at very low concentrations . The results are consistent with a model of cell-cell adhesion in which the HAI is a multivalent, extracellular aggregation factor that is recognized by pallidin molecules on adjacent cells. The HAI would then be analogous to the aggregation factors identified in marine sponges.Aggregation-competent amoebae of the cellular slime mold Polysphondylium pallidum contain a soluble galactose-binding hemagglutinin (1) . This lectin, pallidin, consists of a family of three closely related isolectins, each consisting of different combinations of three subunits of 25-27 kdaltons (2, 3). Considerable evidence indicates that this lectin may be involved in intercellular adhesion ofthe slime mold amoebae : (a) the lectin is present in an active carbohydrate-binding form on the cell surface of differentiated, mutually adhesive amoebae and in a lesser amount on vegetative amoebae (1, 4); (b) high-affinity receptor sites for pallidin are present on the cell surface of differentiated amoebae, as detected by agglutination studies, binding measurements, and cytochemical means (1, 4, 5); (c) high concentrations (>50 mM) of specific saccharide inhibitors of pallidin (lactose or D-galactose) selectively block the cohesiveness of amoebae in swirled suspensions (1) ; (d) the specific lectin antagonists, asialofetuin and immune F'ab, also block the intercellular adhesion ofgyrated amoebae when the assays are carried out in hypertonic media or in the presence of antimetabolites (6, 7). These results suggest that the complementary interaction between pallidin and its receptor on adjoining cells is involved in the mediation ofcell-cell adhesion. Recent genetic experiments have established that the principal

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Inhibition of intercellular adhesion in a cellular slime mould by univalent antibody against a cell-surface lectin

Cited by 39 publications

References 13 publications

Cell adhesion molecules: detection with univalent second antibody.

Cell adhesion molecules: detection with univalent second antibody.

Use of Microarrays as a High-Throughput Platform for Label-Free Biosensing

Identification and purification of an endogenous receptor for the lectin pallidin from Polysphondylium pallidum.

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