Max-Planck Institute for Polymer Research , Ackermannweg 10 , 55128 Mainz , Germany.
Institute of Inorganic Chemistry I , University of Ulm , Albert-Einstein-Allee 11 , 89081 Ulm , Germany.
Bioconjug Chem. 2018 Aug 15;29(8):2665-2670. doi: 10.1021/acs.bioconjchem.8b00358. Epub 2018 Jul 24.
The development of small protein tags that exhibit bioorthogonality, bond stability, and reversibility, as well as biocompatibility, holds great promise for applications in cellular environments enabling controlled drug delivery or for the construction of dynamic protein complexes in biological environments. Herein, we report the first application of dynamic covalent chemistry both for purification and for reversible assembly of protein conjugates using interactions of boronic acid with diols and salicylhydroxamates. Incorporation of the boronic acid (BA) tag was performed in a site-selective fashion by applying disulfide rebridging strategy. As an example, a model protein enzyme (lysozyme) was modified with the BA tag and purified using carbohydrate-based column chromatography. Subsequent dynamic covalent "click-like" bioconjugation with a salicylhydroxamate modified fluorescent dye (BODIPY FL) was accomplished while retaining its original enzymatic activity.
小分子蛋白标签的发展具有生物正交性、键稳定性和可逆性以及生物相容性，有望在细胞环境中实现控制药物释放的应用，或者在生物环境中构建动态蛋白质复合物。在此，我们首次应用动态共价化学，通过硼酸与二醇和水杨羟肟酸的相互作用，用于蛋白质缀合物的纯化和可逆组装。通过二硫键重桥接策略，以选择性方式进行硼酸（BA）标签的掺入。例如，通过 BA 标签对模型蛋白酶（溶菌酶）进行修饰，并使用基于碳水化合物的柱色谱进行纯化。随后，与经过水杨羟肟酸修饰的荧光染料（BODIPY FL）进行动态共价“点击样”生物偶联，同时保留其原始的酶活性。
