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Concanavalin A as A Quantitative and Ultrastructural Probe for Normal and Neoplastic Cell Surfaces

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Concanavalin A

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 55))

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 ultra-structural labeling with fluorescent- or ferritin-Con A. Con A agglutinates confluent-SV3T3 and 3T3 cells at midpoints of 20–60 and > 1,500–2,000 μg/ml, respectively, and sparse cells at 5–15 and 1,2001,500 μg/ml, respectively. Quantitative binding of 125I-Con A at 4°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°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°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°C, washed and then shifted to 20–37°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 sub-plasma 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.

Supported by N.C.I. contract CB-33879 from the Tumor Immunology Program and grant CA-75122, NSF grant P4B1719 from the Human Cell Biology Program and a grant from the Cancer Research Institute, Inc.

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  1. The Salk Institute for Biological Studies, San Diego, California, USA

    Garth L. Nicolson

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  1. Department of Physiology and Biophysics, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA

    Tushar K. Chowdhury  & A. Kurt Weiss  & 

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© 1975 Plenum Press, New York

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Nicolson, G.L. (1975). Concanavalin A as A Quantitative and Ultrastructural Probe for Normal and Neoplastic Cell Surfaces. In: Chowdhury, T.K., Weiss, A.K. (eds) Concanavalin A. Advances in Experimental Medicine and Biology, vol 55. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-0949-9_8

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  • DOI: https://doi.org/10.1007/978-1-4684-0949-9_8

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