Crystallographic characterization of a tri-Asp metal-binding site at the three-fold symmetry axis of LarE
Crystallographic characterization of a tri-Asp metal-binding site at the three-fold symmetry axis of LarE
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DOI：
10.1038/s41598-020-62847-6
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发表时间：
2020-04-02
影响因子：
4.6
通讯作者：
Hu, Jian
中科院分区：
文献类型：
--
作者：
Fellner, Matthias;Huizenga, Kamren G.;Hu, Jian
详细的晶体学表征的三天冬氨酸金属结合位点先前确定的三倍对称轴的六聚体酶，LarE从植物乳杆菌，进行。通过筛选一系列单价、二价和三价金属离子，我们证明了该金属结合位点化学计量地结合Ca 2+、Mn 2+、Fe 2 +/Fe 3+、Co 2+、Ni 2+、Cu 2+、Zn 2+和Cd 2+，但不结合单价金属离子Cr 3+、Mg 2+、Y 3+、Sr 2+或Ba 2+。广泛的数据库搜索导致在其他蛋白质中只有13个类似的金属结合位点，这表明三天冬氨酸结构的稀有性，这允许将这种选择性多价金属离子结合位点工程化到靶大分子中用于结构和生物物理表征。
Detailed crystallographic characterization of a tri-aspartate metal-binding site previously identified on the three-fold symmetry axis of a hexameric enzyme, LarE from Lactobacillus plantarum, was conducted. By screening an array of monovalent, divalent, and trivalent metal ions, we demonstrated that this metal binding site stoichiometrically binds Ca2+, Mn2+, Fe2+/Fe3+, Co2+, Ni2+, Cu2+, Zn2+, and Cd2+, but not monovalent metal ions, Cr3+, Mg2+, Y3+, Sr2+ or Ba2+. Extensive database searches resulted in only 13 similar metal binding sites in other proteins, indicative of the rareness of tri-aspartate architectures, which allows for engineering such a selective multivalent metal ion binding site into target macromolecules for structural and biophysical characterization.