Reversible oriented surface immobilization of functional proteins on oxide surfaces
Reversible oriented surface immobilization of functional proteins on oxide surfaces
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DOI：
10.1021/ac9700033
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发表时间：
1997-06-01
影响因子：
7.4
通讯作者：
Vogel, H
中科院分区：
文献类型：
--
作者：
Schmid, EL;Keller, TA;Vogel, H
在固体表面上以功能活性形式可逆定向固定蛋白质是利用表面敏感技术研究分子相互作用的先决条件。我们展示了一种通常适用于蛋白质附着到氧化物表面的方法。通过硅烷化学将一硝基三乙酸基团作为过渡金属离子的螯合剂与表面共价结合。用六组氨酸延伸修饰的绿色荧光蛋白的可逆结合，用全内反射荧光原位监测。本文所描述的蛋白质在表面上的可逆定向固定化为蛋白质和受体-配体相互作用的结构研究开辟了新的途径。
Reversible and oriented immobilization of proteins in a functional active form on solid surfaces is a prerequisite for the investigation of molecular interactions by surface-sensitive techniques. We demonstrate a method generally applicable for the attachment of proteins to oxide surfaces. A nitrilotriacetic acid group sewing as a chelator for transition metal ions was covalently bound to the surface via silane chemistry. Reversible binding of the green fluorescent protein, modified with a hexahistidine extension, was monitored in situ using total internal reflection fluorescence. The association constant and kinetic parameters of the binding process were determined, The reversible, directed immobilization of proteins on surfaces as described here opens new ways for structural investigation of proteins and receptor-ligand interactions.