Molecular modelling studies of lysozyme catalysed hydrolysis of synthetic substrates.
Kinetic data for the lysozyme catalysed hydrolysis of aryl chitooligosides were surveyed. Both electron-donating and electron-withdrawing substituents on the departing aryl aglycones enhance the rate of hydrolyses. The parallel pH-rate profiles implicate that identical catalytic residues are involved in the hydrolytic fission of the glycosyl-aryloxy bond of these two groups of synthetic substrates. Molecular modelling studies of lysozyme complexes with aryl diN-acetyl chitobiosides and their intermediates were performed. The two synthetic substrates bearing aryl aglycones with opposite electronic effects bind to the active site of lysozyme in different conformations. Based on the energetic and geometric considerations, the oxocarbonium ion whose pyranose ring D in a sofa conformation is the most plausible reaction intermediate for the lysozyme catalysed hydrolysis of the synthetic substrates. The modelling study also suggests that considerable conformational changes of both the lysozyme binding site and the chitobiosyl group accompany the formation of the glycosyl enzyme intermediate. In particular, the chitobiosyl group undergoes a dislocation of the pyranose ring C from the subsite C and a constraint of the pyranose ring D to form a boat conformer.
Tsai
C S
CS
Department of Chemistry, Carleton University, Ottawa, Ontario, Canada.
D000818:Animals; D002642:Chick Embryo; D006868:Hydrolysis; D008958:Models, Molecular; D009113:Muramidase / Q000378:metabolism*; D013379:Substrate Specificity
D009113:Muramidase