Contributions by ionic and steric features of ligands to their binding with phosphorylcholine-specific immunoglobulin IgA H-8 as determined by fluorescence spectroscopy.
The murine myeloma IgA H-8 Fab' fragment which exhibits a binding specificity for phosphorylcholine was assayed for its ability to bind with a number of charged ligands. Monitoring of the ligand-induced changes of protein fluorescence provided a fast and accurate method of determining the equilibrium binding constants. The binding data along with fluorescence spectral properties of the protein permitted an assessment of the relative importance of some binding parameters as well as an evaluation of certain ionic and steric contributions made by ligands exhibiting significant binding affinity for the antibody fragment. Among the conclusions reached is that the dielectric of the binding site microenvironment is important in determining the strength of binding and that hydrophobic groups surrounding a quaternary cationic ligand are important in creating an appropriate binding site of low dielectric value.
D000818:Animals; D002151:Calorimetry; D002794:Choline / Q000031:analogs & derivatives; D007070:Immunoglobulin A; D007140:Immunoglobulin Fab Fragments; D007700:Kinetics; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008968:Molecular Conformation; D009374:Neoplasms, Experimental / Q000276:immunology; D010767:Phosphorylcholine; D010954:Plasmacytoma / Q000276:immunology; D011487:Protein Conformation; D000644:Quaternary Ammonium Compounds; D013050:Spectrometry, Fluorescence
D007070:Immunoglobulin A; D007140:Immunoglobulin Fab Fragments; D008024:Ligands; D000644:Quaternary Ammonium Compounds; D010767:Phosphorylcholine; D002794:Choline