Cell surface interactions with concanavalin A. Location of bound radiolabeled lectin

P. G. Phillips, P. Furmanski, M. Lubin

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

We have developed two improved methods: (1) a procedure for coupling 125Iodine to ConcanavalinA (ConA) that yields intensely labeled and fully active lectin; (2) a procedure that allows studies of lectin binding to be carried out with a minimum of non-specific binding to reaction vessels. We found that BALB/c 3T3 cells, SV3T3 cells, and human red blood cells have 1.3 × 107, 1.5 × 107, and 2.2 × 106 ConA binding sites/cell. More than 99.5% of the radioactivity in the samples counted was associated with the cells; background radioactivity, in the absence of cells, was negligible. We also found that although α-methylmannopyranoside (α-MM) prevented almost all of the ConA from binding to cells, when ConA had first been allowed to bind, α-MM removed only 60 to 80% of the bound ConA. In addition, even after the removal of a portion of bound lectin by α-MM, most, if not all, of the remaining cell-associated ConA was coupled to the plasma membrane.

Original languageEnglish (US)
Pages (from-to)301-308
Number of pages8
JournalExperimental cell research
Volume86
Issue number2
DOIs
StatePublished - Jan 1 1974
Externally publishedYes

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Concanavalin A
Lectins
Cell Communication
Methylmannosides
Radioactivity
BALB 3T3 Cells
Erythrocytes
Binding Sites
Cell Membrane

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

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abstract = "We have developed two improved methods: (1) a procedure for coupling 125Iodine to ConcanavalinA (ConA) that yields intensely labeled and fully active lectin; (2) a procedure that allows studies of lectin binding to be carried out with a minimum of non-specific binding to reaction vessels. We found that BALB/c 3T3 cells, SV3T3 cells, and human red blood cells have 1.3 × 107, 1.5 × 107, and 2.2 × 106 ConA binding sites/cell. More than 99.5{\%} of the radioactivity in the samples counted was associated with the cells; background radioactivity, in the absence of cells, was negligible. We also found that although α-methylmannopyranoside (α-MM) prevented almost all of the ConA from binding to cells, when ConA had first been allowed to bind, α-MM removed only 60 to 80{\%} of the bound ConA. In addition, even after the removal of a portion of bound lectin by α-MM, most, if not all, of the remaining cell-associated ConA was coupled to the plasma membrane.",
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Cell surface interactions with concanavalin A. Location of bound radiolabeled lectin. / Phillips, P. G.; Furmanski, P.; Lubin, M.

In: Experimental cell research, Vol. 86, No. 2, 01.01.1974, p. 301-308.

Research output: Contribution to journalArticle

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