Protein supramolecular complex formation by site-specific avidin-biotin interactions
Protein supramolecular complex formation by site-specific avidin-biotin interactions
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DOI：
10.1039/c2ob26625c
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发表时间：
2013-01-01
影响因子：
3.2
通讯作者：
Kamiya, Noriho
中科院分区：
文献类型：
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作者：
Mori, Yutaro;Wakabayashi, Rie;Kamiya, Noriho
通过基于分子自组装的自下而上的方法，在纳米尺度上精确积累蛋白质功能以制造先进的生物材料已经成为可能。亲和素-生物素相互作用被广泛应用于功能蛋白质自组装体的设计。在此，我们评估了蛋白质构件之间的抗生物素蛋白-生物素相互作用的空间排列如何影响蛋白质超分子复合物（PSC）的形成。对称蛋白质支架，细菌碱性磷酸酶（AP），与专门设计的生物素化底物的酶位点特异性内部标记显示，AP上的生物素化位点的精确定位和底物的接头灵活性是在链霉亲和素（SA）的存在下PSC生长的关键因素。在酶联免疫吸附试验中证明了由AP和SA组成的PSC的潜在诊断应用。
The precise accumulation of protein functions on a nanoscale to fabricate advanced biomaterials has become possible by a bottom-up approach based on molecular self-assembly. The avidin-biotin interaction is widely employed in the design of functional protein self-assemblies. Herein we assessed how the spatial arrangement of the avidin-biotin interaction between protein building blocks affects the formation of a protein supramolecular complex (PSC). The enzymatic site-specific internal labeling of a symmetric protein scaffold, bacterial alkaline phosphatase (AP), with specifically designed biotinylation substrates revealed that the precise positioning of the biotinylation sites on AP and the linker flexibility of the substrate are critical factors for the growth of PSCs in the presence of streptavidin (SA). A potential diagnostic application of the PSCs comprised of AP and SA was demonstrated in an enzyme-linked immunosorbent assay.