Assessing the influence of adsorbed-state conformation on the bioactivity of adsorbed enzyme layers.
Assessing the influence of adsorbed-state conformation on the bioactivity of adsorbed enzyme layers.
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DOI：
10.1021/la900799m
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发表时间：
2009-12-15
期刊：
影响因子：
--
通讯作者：
Latour RA
中科院分区：
文献类型：
--
作者：
Fears KP;Latour RA
使用固定化酶的系统对于广泛的工业和医疗应用是有吸引力的，因为它们允许制造具有高度特异性功能的稳定的、可重复使用的底物。这些系统的性能在很大程度上取决于吸附的酶的取向和构象。为了研究这些关系，我们已经开发和应用的方法来定量评估二级结构的平面表面上吸附的酶层，使用圆二色性（CD）光谱和评估其生物活性，使用比色测定。这些结合的测量提供了分子水平的见解，无论是观察到的吸附酶的生物活性的变化是由于吸附方向的酶或吸附诱导的变化，其构象。利用这种方法，我们研究了溶菌酶（HEWL），木聚糖酶（XYL），和葡萄糖氧化酶（GOx）的OH-，CH 3-，NH 2-，和COOH-封端的烷基乙醇自组装单分子膜（SAM）表面的吸附行为。的小酶，HEWL和XYL的生物活性，有显着的变化之间的不同SAM表面，尽管它们的结构稳定性，突出的吸附取向的作用，生物活性。相比之下，GOx，这是一个更大的酶，吸附后，其结构和生物活性表现出广泛的变化，吸附诱导的构象变化，实际上提高其生物活性。这些结果提供了新的见解，在分子水平上的蛋白质表面相互作用，并表明，吸附可以促进或抑制生物活性取决于表面化学如何影响的方向和构象状态的酶的表面上。
Systems using immobilized enzymes are attractive for a wide range of industrial and medical applications because they allow for the fabrication of stable, reusable substrates with highly specific functionality. The performance of these systems is greatly dependent upon the orientation and conformation of the adsorbed enzymes. To investigate these relationships, we have developed and applied methods to quantitatively assess the secondary structure of adsorbed enzyme layers on planar surfaces using circular dichroism (CD) spectroscopy and evaluate their bioactivity using colorimetric assays. These combined measurements provide molecular-level insights regarding whether observed changes in adsorbed enzyme bioactivity are due to the adsorbed orientation of an enzyme or adsorption-induced changes in its conformation. Using this approach, we investigated the adsorption behavior of lysozyme (HEWL), xylanase (XYL), and glucose oxidase (GOx) on OH-, CH3-, NH2-, and COOH-terminated alkanethiol self-assembled monolayer (SAM) surfaces. The bioactivities of the small enzymes, HEWL and XYL, had pronounced variations between the different SAM surfaces despite their structural stability, highlighting the role of adsorbed orientation on bioactivity. In contrast, GOx, which is a much larger enzyme, exhibited wide variations in both its structure and bioactivity after adsorption, with adsorption-induced conformational changes actually enhancing its bioactivity. These results provide new insights into protein-surface interactions at the molecular level and demonstrate that adsorption can either promote or inhibit bioactivity depending on how the surface chemistry influences the orientation and conformational state of the enzyme on the surface.
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影响因子：
3.9
作者：
Fears, Kenan P.;Creager, Stephen E.;Latour, Robert A.
通讯作者：
Latour, Robert A.
影响因子：
3
作者：
Godjevargova, T;Vasileva, N;Letskovska, Z
通讯作者：
Letskovska, Z
影响因子：
--
作者：
LAYNE, E
通讯作者：
LAYNE, E
影响因子：
3.9
作者：
Agashe, M;Raut, V;Latour, RA
通讯作者：
Latour, RA
影响因子：
3.5
作者：
Lei, Chenghong;Soares, Thereza A.;Ackerman, Eric J.
通讯作者：
Ackerman, Eric J.