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Analysis of murine interferon‐γ binding to its receptor on intact cells and solubilized membranes Identification of an 80 kDa receptor
Author(s) -
Cofano F.,
Landolfo S.,
Appella ]E.,
Ullrich S.J.
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)80475-2
Subject(s) - chaps , receptor , molecular mass , membrane , cell surface receptor , solubilization , microbiology and biotechnology , chemistry , biochemistry , cell culture , biology , enzyme , genetics
The receptor for murine‐interferon‐γ (Mu‐IFN‐γ) has been characterized for its molecular size and equilibrium binding constant on a thymoma cell line, EL‐4. Binding of 125 I‐IFN‐γ to intact cells and their solubilized membranes has shown a single class of receptor with K d values of 1.9 × 10 −9 M and 1.3 × 10 −8 M, respectively. It was shown that solubilization of the Mu‐IFN‐γ receptor with a Zwitterionic detergent (Chaps) preserves its binding activity. A direct comparison of the molecular mass of the Mu‐IFN‐γ receptor on intact cells versus detergent‐solubilized membranes was performed using a radiolabeled photoactivated crosslinking reagent and direct hybridization with 125 I‐labeled IFN‐γ on Western blots of solubilized receptor. The results indicate that both types of receptors have an identical molecular mass of ∼80 kDa.