Atomic-Scale View of Protein-PEG Interactions that Redirect the Thermal Unfolding Pathway of PEGylated Human Galectin-3.
Atomic-Scale View of Protein-PEG Interactions that Redirect the Thermal Unfolding Pathway of PEGylated Human Galectin-3.
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DOI：
10.1002/anie.202203784
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发表时间：
2022-10-04
影响因子：
16.6
通讯作者：
Eddy, Matthew T.
中科院分区：
文献类型：
--
作者：
Pritzlaff, Amanda;Ferre, Guillaume;Mulry, Emma;Lin, Ling;Pour, Niloofar Gopal;Savin, Daniel A.;Harris, Michael E.;Eddy, Matthew T.
聚乙二醇化是一种很有前途的方法，可以解决应用生物制剂的核心挑战：即在人体的苛刻环境中缺乏蛋白质稳定性。由于缺乏对蛋白质-PEG相互作用的原子水平理解，阻碍了更广泛的应用，从而阻止了具有预测性质的缀合物的设计。我们部署了一种综合的结构和生物物理方法，以解决这一关键挑战与聚乙二醇化的碳水化合物识别结构域的人半乳糖凝集素-3（Gal 3C），凝集素细胞粘附和潜在的生物。PEG化显著增加了Gal 3C的热稳定性，形成稳定的中间体并重新定向其解折叠途径。NMR揭示的结构细节指出了带电氨基酸促进PEG定位的重要作用。替换这些残基改变了蛋白质-PEG界面和热解折叠行为，提供了合理设计缀合物同时保留PEG化益处的策略。人半乳糖凝集素-3的碳水化合物识别结构域的PEG化显著地重定向蛋白质热解折叠途径。NMR光谱显示PEG定位于蛋白质表面，与“裹尸布”分子模型一致。PEG的本地化和相应的热去折叠配置文件的微妙变化，实现了通过更换赖氨酸附近的蛋白质-PEG界面。
PEGylation is a promising approach to address the central challenge of applying biologics: i.e. lack of protein stability in the demanding environment of the human body. Wider application is hindered by lack of atomic level understanding of protein–PEG interactions, preventing design of conjugates with predicted properties. We deployed an integrative structural and biophysical approach to address this critical challenge with the PEGylated carbohydrate recognition domain of human galectin-3 (Gal3C), a lectin essential for cell adhesion and potential biologic. PEGylation dramatically increased Gal3C thermal stability, forming a stable intermediate and redirecting its unfolding pathway. Structural details revealed by NMR pointed to an important role of PEG localization facilitated by charged amino acids. Replacing these residues altered the protein–PEG interface and thermal unfolding behavior, providing a strategy to rationally design conjugates while preserving PEGylation benefits. PEGylation of the carbohydrate recognition domain of human Galectin-3 dramatically redirected the protein thermal unfolding pathway. NMR spectroscopy showed PEG localized to the protein surface consistent with a ‘shroud’ molecular model. Subtle changes in the PEG localization and corresponding thermal unfolding profiles were realized by replacing lysines near the protein-PEG interface.
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影响因子：
4.7
作者：
Dhalluin, C;Ross, A;Senn, H
通讯作者：
Senn, H
影响因子：
16.6
作者：
Lee, Cheng-Chung;Maestre-Reyna, Manuel;Hsu, Kai-Cheng;Wang, Hao-Ching;Liu, Chia-I;Jeng, Wen-Yih;Lin, Li-Ling;Wood, Richard;Chou, Chia-Cheng;Yang, Jinn-Moon;Wang, Andrew H. -J.
通讯作者：
Wang, Andrew H. -J.
影响因子：
3.7
作者：
Mirandola L;Yu Y;Chui K;Jenkins MR;Cobos E;John CM;Chiriva-Internati M
通讯作者：
Chiriva-Internati M
影响因子：
21.8
作者：
Krall, Nikolaus;da Cruz, Filipa P.;Bernardes, Goncalo J. L.
通讯作者：
Bernardes, Goncalo J. L.
影响因子：
4.6
作者：
Grigoletto, Antonella;Mero, Anna;Pasut, Gianfranco
通讯作者：
Pasut, Gianfranco