Residue-level elucidation of the ligand-induced protein binding on phenyl-argarose microspheres by NMR hydrogen/deuterium exchange technique
Residue-level elucidation of the ligand-induced protein binding on phenyl-argarose microspheres by NMR hydrogen/deuterium exchange technique
复制标题
通过 NMR 氢/氘交换技术对苯基琼脂糖微球上配体诱导的蛋白质结合进行残留水平阐明
DOI：
10.1039/c2sm25117e
复制
发表时间：
2012-01-01
期刊：
影响因子：
3.4
通讯作者：
Su, Zhi-Guo
中科院分区：
文献类型：
--
作者：
Hao, Dong-Xia;Sandstrom, Corine;Su, Zhi-Guo
蛋白质-配体在液-固界面上的相互作用决定了功能生物材料的设计。然而，准确的残基细节的配体诱导的蛋白质结合和展开的界面上仍然未知的蛋白质结构表征的当前合奏。在这里，氢/氘（H/D）的方法加上NMR TOCSY光谱分析和溶剂可及表面积（SASA）的设计，使残留水平的理解吸附在苯基配体修饰的表面的溶菌酶。结果表明，溶菌酶分子在苯基琼脂糖微球上的结合位点和去折叠具有明显的配体密度依赖性和蛋白质覆盖度依赖性。无论是增加配体密度还是降低吸附覆盖度，都会导致更多的结合位点和蛋白质分子的解折叠。随着多点吸附的增强，蛋白质分子由端对侧吸附转变为侧对侧吸附。最后，利用Molecular Dock模拟方法对NMR确定的结合位点进行了能量排序。它证实，这种NMR方法代表了一种可靠的途径，在与生物材料的蛋白质相互作用过程中，在硅片上丰富的残基水平的结构信息。
Protein-ligand interactions on liquid-solid interfaces governed the design of functional biomaterials. However, accurate residue details of ligand induced protein binding and unfolding on an interface were still unknown by the current ensemble of protein structure characterizations. Here, a hydrogen/deuterium (H/D) approach coupled with analysis of NMR TOCSY spectra and the solvent accessible surface area (SASA) was designed to enable residue level understanding of lysozyme adsorbed at a phenyl-ligand modified surface. Results showed that the binding sites and unfolding of lysozyme molecules on phenyl-agarose microspheres demonstrated significant ligand-density dependence and protein-coverage dependence. Either increasing ligand density or decreasing adsorption coverage would lead to more binding sites and unfolding of the protein molecules. With the multipoint adsorption strengthening, the protein molecule changed from lying end-on to side-on. Finally, Molecular Dock simulation was utilized to evaluate the NMR determined binding sites based on energy ranking of the binding. It confirmed that this NMR approach represents a reliable route to in silico abundant residue-level structural information during protein interaction with biomaterials.