Automated hydrodynamic modelling of a complex between a human IgE fragment (Fc epsilon 3-4) and the IgE high affinity receptor Fc epsilon RI alpha-chain.
The binding of IgE to its high affinity receptor Fc epsilon RI plays an important role in the allergic response. The interaction between soluble Fc epsilon RI alpha-chain (sFc epsilon RI alpha) and Fc epsilon 3-4, a fragment of IgE consisting of the C epsilon 3 and C epsilon 4 heavy chain constant domains, has been studied using analytical ultracentrifugation (Keown et al. this volume). Here we describe the development of a simple automated hydrodynamic modelling technique and its application to this interaction. This procedure utilises sphere models of the two molecules and performs an automated systematic translational search of sFc epsilon RI alpha relative to Fc epsilon 3-4. The result of this is the generation of 40,359 individual models of how the receptor can be placed relative to Fc epsilon 3-4. These are then assessed for consistency by comparing the sedimentation coefficients generated for the models to the experimentally determined sedimentation coefficients, and are displayed graphically to show allowed and disallowed complexes. From this analysis, it is clear that the complex between sFc epsilon RI alpha and Fc epsilon 3-4 is compact, with the most elongated models being excluded. In addition, sFc epsilon RI alpha appears not to interact with the C-terminal end of Fc epsilon 3-4, and probably binds either to the sides or face, observations which are consistent with other experimental data on the Fc epsilon RI alpha/IgE interaction. Automated hydrodynamic modelling also has the potential to be used for other interactions, providing a simple way of looking at a large number of models, and making rigorous studies of interacting components more feasible.
Beavil
A J
AJ
Randall Institute, King's College London, United Kingdom.
Beavil
R L
RL
D001331:Automation; D001665:Binding Sites; D006801:Humans; D007073:Immunoglobulin E / Q000737:chemistry* / Q000378:metabolism; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry* / Q000378:metabolism; D007139:Immunoglobulin epsilon-Chains / Q000737:chemistry* / Q000378:metabolism; D046911:Macromolecular Substances; D011487:Protein Conformation; D017455:Receptors, IgE / Q000737:chemistry* / Q000378:metabolism; D014461:Ultracentrifugation / Q000379:methods
D007141:Immunoglobulin Fc Fragments; D007139:Immunoglobulin epsilon-Chains; D046911:Macromolecular Substances; D017455:Receptors, IgE; D007073:Immunoglobulin E