Thermodynamics of intersubunit interactions in cholera toxin upon binding to the oligosaccharide portion of its cell surface receptor, ganglioside GM1.
The binding and the energetics of the interaction of cholera toxin with the oligosaccharide portion of ganglioside GM1 (oligo-GM1), the toxin cell surface receptor, have been studied by high-sensitivity isothermal titration calorimetry and differential scanning calorimetry. Previously, we have shown that the association of cholera toxin to ganglioside GM1 enhances the cooperative interactions between subunits in the B-subunit pentamer [Goins, B., & Freire, E. (1988) Biochemistry 27, 2046-2052]. New experiments presented in this paper reveal that the oligosaccharide portion of the receptor is by itself able to enhance the intersubunit cooperative interactions within the B pentamer. This effect is seen in the protein unfolding transition as a shift from independent unfolding of the B promoters toward a cooperative unfolding. To identify the origin of this effect, the binding of cholera toxin to oligo-GM1 has been measured calorimetrically under isothermal conditions. The binding curve at 37 degrees C is sigmoidal, indicating cooperative binding. The binding data can be described in terms of a nearest-neighbor cooperative interaction binding model. In terms of this model, the association of a oligo-GM1 molecule to a B protomer affects the association to adjacent B promoters within the pentameric ring. The measured intrinsic binding enthalpy per protomer is -22 kcal/mol and the cooperative interaction enthalpy -11 kcal/mol. The intrinsic binding constant determined calorimetrically is 1.05 x 10(6) M-1 at 37 degrees C and the cooperative Gibbs free energy equal to -850 cal/mol.(ABSTRACT TRUNCATED AT 250 WORDS)
Schön
A
A
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
Freire
E
E
D001665:Binding Sites; D001667:Binding, Competitive; D002151:Calorimetry; D002152:Calorimetry, Differential Scanning; D002772:Cholera Toxin / Q000032:analysis*; D004735:Energy Transfer; D005677:G(M1) Ganglioside / Q000032:analysis*; D008565:Membrane Proteins / Q000378:metabolism*; D009844:Oligosaccharides / Q000032:analysis*; D013696:Temperature; D013816:Thermodynamics
D008565:Membrane Proteins; D009844:Oligosaccharides; D005677:G(M1) Ganglioside; D002772:Cholera Toxin