Chemical Strategy for Studying the Antigenic Structures of Disulfide-Containing Proteins: Hen Egg-White Lysozyme as a Model

摘要: A critical approach in studying the immunochemistry of proteins relies on preparation a variety long and overlapping peptides from parent molecule. Disulfide-containing are usually inaccessible to proteolytic attack therefore it is not possible prepare such without rupturing disulfides. Unfortunately, preparations with broken disulfides bear no immunochemical relationship native proteins. These have until recently virtually eluded investigation. break deadlock, we introduced novel cleavage for obtaining protein disulfide bonds. Reversible citraconylation amino groups induced conformational changes that were sufficient render completely accessible tryptic hydrolysis at arginyl Complete lysyl bonds may be effected, if desired, after deprotection which proceeds quantitatively. by this obtained lysozyme accounted almost all (907%) immune reaction lysozyme. From these numerous chemical derivatives lysozyme, was derive location antigenic sites Two then accurately delineated. One achieved organic synthesis 9 corresponding various overlaps around 6–127. delineation second site (around 64–80 76–94), devised (Atassi, Lee Pai, 1976, Biochim. Biophys. Acta, 427, 745) an entirely “Surface-Simulation” synthetic approach. this, conformationally-adjacent residues constructing linked directly via peptide into single does exist but simulates surface region it. This unorthodox provides powerful technique final reactive (and perhaps other types binding sites)