Examining polymer-protein biophysical interactions with small-angle x-ray scattering and quartz crystal microbalance with dissipation.
Examining polymer-protein biophysical interactions with small-angle x-ray scattering and quartz crystal microbalance with dissipation.
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DOI：
10.1002/jbm.a.37479
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发表时间：
2023-04
影响因子：
4.9
通讯作者：
Gormley, Adam J.
中科院分区：
文献类型：
--
作者：
Upadhya, Rahul;Di Mare, Elena;Tamasi, Matthew J.;Kosuri, Shashank;Murthy, N. Sanjeeva;Gormley, Adam J.
关键词：
聚合物-蛋白质杂化材料可用于改善蛋白质在变性环境中的溶解度和稳定性。虽然之前的工作使用了机器人学和主动机器学习来为新的设计提供信息，但需要进一步的生物物理信息来确定结构功能行为。在这里，我们展示了串联小角X射线散射(SAXS)和石英晶体微天平(QCMD)实验的价值，以揭示详细的聚合物-蛋白质与辣根过氧化物酶(HRP)的相互作用。特别令人感兴趣的是在热应力下聚合物-蛋白质复合体的形成过程，其中SAXS监测溶液中的形成，而QCMD在界面上跟踪这些动力学。在变性条件下，SAXS测量的蛋白质的旋转半径(Rg)在聚合物存在的情况下没有显著变化，但在QCMD数据中观察到了厚度和耗散的变化。利用具有和不具有热应力的SAXS数据来创建潜在络合物和变性酶的珠状模型，每个模型拟合提供了对相互作用程度的洞察。此外，QCMD数据表明，HRP在低浓度下通过表面吸附而发生变形，表现为较长的吸附时间和较小的频移。相反，热应激和高度非活性的HRP具有更快的吸附动力学。SAXS和QCMD的结合为蛋白质和聚合物之间相互作用的生物物理表征提供了一个框架，这可能有助于设计聚合物-蛋白质杂化材料。
Polymer-protein hybrids can be deployed to improve protein solubility and stability in denaturing environments. While previous work used robotics and active machine learning to inform new designs, further biophysical information is required to ascertain structure–function behavior. Here, we show the value of tandem small-angle x-ray scattering (SAXS) and quartz crystal microbalance with dissipation (QCMD) experiments to reveal detailed polymer-protein interactions with horseradish peroxidase (HRP) as a test case. Of particular interest was the process of polymer-protein complex formation under thermal stress whereby SAXS monitors formation in solution while QCMD follows these dynamics at an interface. The radius of gyration (Rg) of the protein as measured by SAXS does not change significantly in the presence of polymer under denaturing conditions, but thickness and dissipation changes were observed in QCMD data. SAXS data with and without thermal stress were utilized to create bead models of the potential complexes and denatured enzyme, and each model fit provided insight into the degree of interactions. Additionally, QCMD data demonstrated that HRP deforms by spreading upon surface adsorption at low concentration as shown by longer adsorption times and smaller frequency shifts. In contrast, thermally stressed and highly inactive HRP had faster adsorption kinetics. The combination of SAXS and QCMD serves as a framework for biophysical characterization of interactions between proteins and polymers which could be useful in designing polymer-protein hybrids.
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DOI：
10.1073/pnas.95.21.12271
发表时间：
1998-10-13
影响因子：
11.1
作者：
Höök, F;Rodahl, M;Brzezinski, P
通讯作者：
Brzezinski, P
影响因子：
10
作者：
通讯作者：
--
影响因子：
3.5
作者：
Cha, Hyung Jin;Jang, Do Soo;Choi, Kwan Yong
通讯作者：
Choi, Kwan Yong
影响因子：
4.6
作者：
Kulakova, Alina;Indrakumar, Sowmya;Harris, Pernille
通讯作者：
Harris, Pernille
DOI：
10.1002/anie.201711044
发表时间：
2018-02-05
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Gormley AJ;Yeow J;Ng G;Conway Ó;Boyer C;Chapman R
通讯作者：
Chapman R