Nakano Y, Kawauchi S, Komiyama J, Iijima T
Department of Polymer Science, Tokyo Institute of Technology, Japan.
Biochem Int. 1987 Aug;15(2):303-10.
A method to determine intrinsic binding constants of lysozyme with substrate analogues such as N-acetyl-D-glucosamine dimer and trimer is proposed. The method is based on the competitive interaction of an anionic azo dye with substrate analogues for lysozyme. There are two binding sites for substrate analogues and dyes, respectively, on lysozyme. One binding mode of the substrate analogues to subsites D-F on lysozyme was non-competitive, and another binding mode to subsites A-C was competitive with the dye. From the binding constants obtained it is suggested that the binding of the substrate analogues to subsite D on lysozyme is weaker than the binding to the other subsites.
提出了一种测定溶菌酶与底物类似物（如N-乙酰-D-葡萄糖胺二聚体和三聚体）内在结合常数的方法。该方法基于阴离子偶氮染料与底物类似物对溶菌酶的竞争性相互作用。溶菌酶上分别有两个底物类似物和染料的结合位点。底物类似物与溶菌酶上D-F亚位点的一种结合模式是非竞争性的，而与A-C亚位点的另一种结合模式与染料是竞争性的。从获得的结合常数表明，底物类似物与溶菌酶上D亚位点的结合比与其他亚位点的结合弱。
