STOPPED-FLOW FLUORESCENCE STUDIES ON SACCHARIDE BINDING TO LYSOZYME
STOPPED-FLOW FLUORESCENCE STUDIES ON SACCHARIDE BINDING TO LYSOZYME
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DOI：
10.1042/bj1490411
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发表时间：
1975-01-01
影响因子：
4.1
通讯作者：
HALFORD, SE
中科院分区：
文献类型：
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作者：
HALFORD, SE
用快速反应动力学方法研究了N-乙酰-D-氨基葡萄糖β-1-4-连接三聚体与溶菌酶的结合反应。结果发现，从这个反应的荧光差异光谱的离散段扰动在不同的时间点在结合过程中。结果被解释为初始复合物的形成，反应的快速阶段，扰动色氨酸-62的环境，以及初始复合物的后续和较慢的重排扰动色氨酸-108的环境。在pH 4.4时，在结合反应的重排步骤期间发生从天冬氨酸-101释放质子。一个模型的反应（E，酶; L，配体）：（见文章）该配体与溶菌酶的协会可以可视化在三维条件下作为初始复合物的形成跨越顶部的活性位点裂缝，然后潜水运动的配体进入裂缝。
The binding of the β-1-4-linked trimer ofN-acetyl-D-glucosamine to hen egg-white lysozyme was studied by rapid-reaction-kinetic methods with tryptophyl fluorescence observation of the transients. It was found that discrete segments of the fluorescence-difference spectrum from this reaction were perturbed at different time-points during the binding process. The results were interpretated as the formation of the initial complex, the fast phase of the reaction, perturbing the environment of tryptophan-62 and a subsequent and slower rearrangement of the initial complex perturbing the environment of tryptophan-108. At pH 4.4, the release of protons from aspartate-101 occurred during the rearrangement step of the binding reaction. A model for the reaction is presented (E, enzyme; L, ligand): (see article) The association of this ligand with lysozyme may be visualized in three-dimensional terms as initial complex-formation across the top of the active-site cleft followed by a diving motion of the ligand into the cleft.