Autocatalytic association of proteins by covalent bond formation: a Bio Molecular Welding toolbox derived from a bacterial adhesin.
Autocatalytic association of proteins by covalent bond formation: a Bio Molecular Welding toolbox derived from a bacterial adhesin.
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DOI：
10.1038/srep43564
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发表时间：
2017-03-02
影响因子：
4.6
通讯作者：
Di Guilmi AM
中科院分区：
文献类型：
--
作者：
Bonnet J;Cartannaz J;Tourcier G;Contreras-Martel C;Kleman JP;Morlot C;Vernet T;Di Guilmi AM
来自革兰氏阳性细菌的黏附素中不寻常的分子内交联已被用于开发一种适用于生物技术应用的通用工艺。基于RrgA（肺炎链球菌菌毛粘附素）的晶体结构，我们提供了证据，证明两个工程蛋白片段一旦从亲本蛋白分离出来，在体内和体外都能保持高特异性的共价结合能力。我们确定了该配合物组装的最佳条件，并在2 Å处解出了其晶体结构。此外，我们展示了与抗体生产、纳米组装和细胞表面标记相关的生物技术应用，我们将此过程命名为生物分子焊接。
Unusual intramolecular cross-links present in adhesins from Gram-positive bacteria have been used to develop a generic process amenable to biotechnology applications. Based on the crystal structure of RrgA, the Streptococcus pneumoniae pilus adhesin, we provide evidence that two engineered protein fragments retain their ability to associate covalently with high specificity, in vivo and in vitro, once isolated from the parent protein. We determined the optimal conditions for the assembly of the complex and we solved its crystal structure at 2 Å. Furthermore, we demonstrate biotechnological applications related to antibody production, nanoassembly and cell-surface labeling based on this process we named Bio Molecular Welding.