Lysozyme bound to the D1.3 monoclonal antibody retains enzymatic activity in assays using N-acetylglucosamine oligomers as substrate.
An enzymatic assay using Micrococcus lysodeikticus as substrate had shown that hen egg-white lysozyme (HEL), complexed to the specific monoclonal antibody D1.3, was enzymatically inactive. However, a crystallographic study of the HEL-Fab D1.3 complex at 2.8 A resolution showed that Fab D1.3 is bound to the enzyme at a region remote from the catalytic site, and that no significant conformational change had occurred in the bound lysozyme. To resolve this apparent discrepancy, oligomers of N-acetylglucosamine, containing an average of six residues and which should not be sterically hindered by the monoclonal antibody, were used as substrate in enzyme assays. In these assays lysozyme complexed to antibody D1.3 retained enzymatic activity, thus supporting the results of the crystallographic study and indicating no major conformational change or rigidification of its structure.
D000117:Acetylglucosamine; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D009113:Muramidase / Q000276:immunology / Q000378:metabolism*; D011200:Poultry; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D009113:Muramidase; D000117:Acetylglucosamine