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  • Title: Concanavalin A as a quantitative and ultrastructural probe for normal and neoplastic cell surfaces.
    Author: Nicolson GL.
    Journal: Adv Exp Med Biol; 1975; 55():153-72. PubMed ID: 168743.
    Abstract:
    Concanavalin A (Con A) has been popularly used as a cell surface probe for normal and neoplastic cells. Differences between normal fibroblasts and their transformed derivatives were examined using Con A agglutination, quantitative labeling with 125I-Con A and ultrastructural labeling with fluorescent- or ferritin-Con A. Con A agglutinates confluent-SV3T3 and 3T3 cells at midpoints of 20-60 and greater than, 500-2,000 mug/ml, respectively, and sparse cells at 5-15 and 1,200-1,500 mg/ml, respectively. Quantitative binding of 125I-Con A at 4 degrees C (10 or 15 min) with saturating lectin concentrations does not indicate a difference in the number of Con A receptors on sparse or confluent 3T3 and SV3T3 cells similar to many publications, but contrary to Noonan and Burger (1973). Under these conditions of labeling, ferritin-Con A is not internalized, indicating absence of endocytosis. The lateral mobility of Con A receptors and their relative ability to be aggregated on the cell surface by Con A was investigated with fluorescent- and ferritin-Con A. The initial distribution of Con A receptors on 3T3, SV3T3 and MSV3T3 cells under conditions of labeling at low temperature (0-5 degrees C) or to fixed cells was essentially randomly dispersed, but changes quickly to aggregated on SV3T3 and MSV3T3 (but not 3T3) after shifting the temperature to 20-37 degrees C, indicating, in general, a greater relative mobility of Con A receptors on SV3T3 and MSV3T3 cells. The aggregated Con A receptors seem to be directly involved in cell agglutination because they are usually found at the sites of cell-to-cell contact during 10 min agglutination experiments with ferritin-Con A. When confluent-3T3 cells are labeled on monolayer with ferritin-Con A at 0-4 degrees C, washed and then shifted to 20-37 degrees C for 10-15 min prior to in situ embedding, two classes of Con A receptors can be identified. One class appears to have low relative mobility and is associated with the 3T3 cell's extensive subplasma membrane microfilament network, while the other is capable of being aggregated and eventually endocytosed. On confluent-SV3T3 cells, only the latter class of receptors appears to be present, indicating a possible loss of cytoplasmic control over the distribution and mobility of lectin-binding sites on transformed cell surfaces.
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