title: Thermodynamics of the interaction of human immunoglobulin E with its high-affinity receptor Fc epsilon RI.
abstract: We have employed isothermal titration calorimetry (ITC) and circular dichroism (CD) spectroscopy to characterize the binding of soluble fragments of IgE (IgE-Fc and Fc epsilon 3-4) to a soluble fragment of the high-affinity receptor Fc epsilon RI alpha-chain (sFc epsilon RI alpha). The thermodynamic parameters for the interaction of IgE-Fc and Fc epsilon 3-4 with sFc epsilon RI alpha, determined using ITC, confirm the earlier conclusion that the C epsilon 2 domain is not involved in the interaction and that the stoichiometry of both complexes is 1:1. For both IgE-Fc and Fc epsilon 3-4, the value of Delta H degrees is -36.9 +/- 4.6 kcal mol-1 at 37.3 degreesC and Delta Cp degrees is -820 +/- 120 cal mol-1 K-1. The temperature at which DeltaS degrees is zero is 284 +/- 1 K, indicating that the entropy contribution to the thermodynamics of association is unfavorable at physiological temperature. Of particular interest is the large value of Delta Cp degrees. The large surface area of IgE and Fc epsilon RI alpha that is implicated in complex formation from previous mutagenesis studies on the two proteins may account in part for the magnitude of Delta Cp degrees. Additional contributions may arise from hydration within the binding site and changes in tertiary structure of the individual components of the complex. However, the CD spectra of IgE, IgE-Fc, and Fc epsilon 3-4 complexes with sFc epsilon RI alpha are merely the sum of the spectra of their individual components, indicating that the secondary structure of the immunoglobulin domain folds are preserved on complex formation. Thus, any change in tertiary structure must be limited to the relative disposition of the immunoglobulin domains C epsilon 3 and C epsilon 4 in IgE and the two immunoglobulin-like domains in the alpha-chain of Fc epsilon RI.
authors.affiliation: The Randall Institute, King's College London, UK.
authors.forename: M B
authors.initials: MB
authors.lastname: Keown
authors.forename: A J
authors.initials: AJ
authors.lastname: Henry
authors.forename: R
authors.initials: R
authors.lastname: Ghirlando
authors.forename: B J
authors.initials: BJ
authors.lastname: Sutton
authors.forename: H J
authors.initials: HJ
authors.lastname: Gould
mesh_terms: D002151:Calorimetry; D002942:Circular Dichroism; D019281:Dimerization; D006801:Humans; D007073:Immunoglobulin E / Q000737:chemistry* / Q000378:metabolism*; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry / Q000378:metabolism; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000378:metabolism; D017433:Protein Structure, Secondary; D017455:Receptors, IgE / Q000737:chemistry* / Q000378:metabolism*; D013816:Thermodynamics
chemical_list: D007141:Immunoglobulin Fc Fragments; D007143:Immunoglobulin Heavy Chains; D017455:Receptors, IgE; D007073:Immunoglobulin E