NMR studies of protein hydration and TEMPOL accessibility
NMR studies of protein hydration and TEMPOL accessibility
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DOI：
10.1016/s0022-2836(03)00852-0
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发表时间：
2003-09-12
影响因子：
5.6
通讯作者：
Temussi, PA
中科院分区：
文献类型：
--
作者：
Niccolai, N;Spiga, O;Temussi, PA
了解蛋白质表面与外部分子环境相互作用的机制，对于新药和工程蛋白的合理设计具有重要意义。蛋白质表面可接近性正在成为结构生物学的一个新维度，因为核磁共振方法已经发展到跟踪分子，即使是那些与生理配体不同的分子，如何优先接近蛋白质表面的特定区域。蛋清溶菌酶是蛋白质结构和动力学研究的典范，被选择作为研究蛋白质表面可及性的模型系统。结合水和可溶性自旋标记被用来研究这种酶与分子环境的相互作用，包括游离和结合抑制剂（NAG）(3)。在酶活性位点内没有发现紧密结合的水分子，相反，在可溶性顺磁探针TEMPOL的访问下，这是最容易暴露的。从提出的数据集，溶菌酶对水和TEMPOL分子的表面可及性的综合视图得到。(C) 2003 Elsevier Ltd.版权所有。
Understanding the mechanisms of the interaction between a protein surface and its outer molecular environment is of primary relevance for the rational design of new drugs and engineered proteins. Protein surface accessibility is emerging as a new dimension of Structural Biology, since NMR methods have been developed to follow how molecules, even those different from physiological ligands, preferentially approach specific regions of the protein surface. Hen egg-white lysozyme, a paradigmatic example of the state of the art of protein structure and dynamics, has been selected as a model system to study protein surface accessibility. Bound water and soluble spin-labels have been used to investigate the interaction of this enzyme, both free and bound to the inhibitor (NAG)(3), with its molecular environment. No tightly bound water molecules were found inside the enzyme active site, which, conversely, appeared as the most exposed to visits from the soluble paramagnetic probe TEMPOL. From the presented set of data, an integrated view of lysozyme surface accessibility towards water and TEMPOL molecules is obtained. (C) 2003 Elsevier Ltd. All rights reserved.