Resurveying the Tris buffer solution: the specific interaction between tris(hydroxymethyl)aminomethane and lysozyme.
Resurveying the Tris buffer solution: the specific interaction between tris(hydroxymethyl)aminomethane and lysozyme.
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DOI：
10.1016/j.ab.2008.04.006
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发表时间：
2008-07
影响因子：
2.9
通讯作者：
L. Quan;Dengguo Wei;Xiaolu Jiang;Yang Liu;Zhiyu Li;Na Li;K. Li;Feng Liu;L. Lai
中科院分区：
文献类型：
--
作者：
L. Quan;Dengguo Wei;Xiaolu Jiang;Yang Liu;Zhiyu Li;Na Li;K. Li;Feng Liu;L. Lai
通过使用石英晶体微天平 (QCM) 生物传感器对相互作用进行快速筛选分析，首次证明了一种不寻常的现象，即三（羟甲基）氨基甲烷 (Tris) 和溶菌酶 (LZM) 之间的特异性相互作用。在表面等离子共振（SPR）系统中也观察到了这种现象。使用高效亲和层析 (HPAC) 的进一步研究证实了 LZM 和固定化 Tris 之间的这种特异性相互作用，表观解离常数 (KD) 为 6.7×10−5M。进行分子对接以确定 LZM 和连接至结合臂的 Tris 之间可能的结合模式。估计的结合自由能为-6.34kcal mol−1，对应的KD为2.3×10−5M，与实验值相关良好。根据对接模型，Tris的三个羟基与LZM中的Asp52、Glu35和Ala107形成分子间H键。这项研究强调了定量生化研究中缓冲液选择的重要性。对于溶菌酶配体结合研究，当所研究的配体是弱结合剂时，最好避免使用 Tris。
An unusual phenomenon, the specific interaction between tris(hydroxymethyl)aminomethane (Tris) and lysozyme (LZM), was demonstrated for the first time by rapid screen analysis of interactions using a quartz crystal microbalance (QCM) biosensor. This phenomenon was also observed in a surface plasmon resonance (SPR) system. Further study using high-performance affinity chromatography (HPAC) confirmed this specific interaction between LZM and immobilized Tris with an apparent dissociation constant (KD) of 6.7×10−5M. Molecular docking was carried out to identify possible modes of binding between LZM and Tris linked to a binding arm. The estimated binding free energy was −6.34kcal mol−1, corresponding to a KDof 2.3×10−5M, which correlated well with the experimental value. Based on the docking model, the three hydroxyl groups of Tris form intermolecular H bonds with Asp52, Glu35, and Ala107 in LZM. This study reinforces the importance of buffer selection in quantitative biochemical investigations. For a lysozyme ligand binding study, it is better to avoid using Tris when the ligands under study are weak binders.