The interaction of IgE with rat basophilic leukemia cells. II. Quantitative aspects of the binding reaction

Abstract

Previous studies indicated that cultured rat basophilic leukemia cells have surface receptors which bind IgE with high specificity. In this paper we describe some quantitative aspects of the phenomenon. The reaction mechanism appears to consist of a simple reversible binding reaction with k(1) = 9.6 x 10(4) M(-1) sec(-1) and k(-1) 1.6 x 10(-5) sec(-1) at 37 degrees C. The calculated K(A) is therefore 6 x 10(9) M(-1). The activation energy of binding was found to be 7.8 kcal/mol. The number of binding sites/cell varied between 3 x 10(5) to over 1 x 10(6). The binding was insensitive to pH's between 6-8 but at pH 3.0 complete dissociation of bound IgE occurred in approximately 1 min at 0 degrees C leaving the receptors for IgE intact. Ca(++) plus EDTA and Mg(++) plus EDTA produce a fairly marked reduction in binding capacity though these reagents alone produce much smaller effects.