UCIBIO, Departamento de Química, Faculdade de Ciências e Tecnologia, UCIBIO, Universidade NOVA de Lisboa, Campus Caparica, 2829-516, Caparica, Portugal.
LAQV-REQUIMTE, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre 687, 4169-007, Porto, Portugal.
Biotechnol J. 2019 Nov;14(11):e1800559. doi: 10.1002/biot.201800559. Epub 2019 Aug 8.
Affinity-triggered assemblies rely on affinity interactions as the driving force to assemble physically crosslinked networks. WW domains are small hydrophobic proteins binding to proline-rich peptides that are typically produced in the insoluble form. Previous works attempted the biological production of the full WW domain in tandem to generate multivalent components for affinity-triggered hydrogels. In this work, an alternative approach is followed by engineering a 13-mer minimal version of the WW domain that retains the ability to bind to target proline-rich peptides. Both ligand and target peptides are produced chemically and conjugated to multivalent polyethylene glycol, yielding two components. Upon mixing together, they form soft biocompatible affinity-triggered assemblies, stable in stem cell culture media, and display mechanical properties in the same order of magnitude as for those hydrogels formed with the full WW protein in tandem.
基于亲和力的组装依赖于亲和力相互作用作为组装物理交联网络的驱动力。WW 结构域是一种小分子疏水蛋白，能够与富含脯氨酸的肽结合，而富含脯氨酸的肽通常以不溶的形式产生。以前的工作试图通过生物串联生产完整的 WW 结构域来产生用于亲和力触发水凝胶的多价组件。在这项工作中，采用了一种替代方法，通过工程设计保留了与靶标富含脯氨酸肽结合能力的 13 肽最小 WW 结构域版本。配体和靶肽都是通过化学方法合成的，并与多价聚乙二醇连接，得到两种成分。将它们混合在一起，形成柔软的生物相容性亲和力触发组装体，在干细胞培养基中稳定，并表现出与串联使用完整 WW 蛋白形成的水凝胶相同数量级的机械性能。
