Adsorption kinetics, conformation, and mobility of the growth hormone and lysozyme on solid surfaces, studied with TIRF
Adsorption kinetics, conformation, and mobility of the growth hormone and lysozyme on solid surfaces, studied with TIRF
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DOI：
10.1006/jcis.1997.4876
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发表时间：
1997-06-01
影响因子：
9.9
通讯作者：
Hlady, V
中科院分区：
文献类型：
--
作者：
Buijs, J;Hlady, V
利用全内反射荧光（TIRF）和色氨酸内源荧光监测技术研究了重组人生长激素和溶菌酶与固体表面的相互作用。监测由吸附的蛋白质发射的荧光的强度、光谱、猝灭和偏振，并将其与吸附动力学、蛋白质构象和荧光团旋转迁移率相关。为了研究静电和疏水相互作用对吸附过程的影响，使用了三种不同电荷和疏水性的吸附剂表面。化学表面基团是硅烷醇、甲基和季胺。结果表明，hGH的吸附主要是由疏水相互作用。离子强度对溶菌酶的吸附有很大的影响。这种效应可能是由溶液中的离子强度依赖性构象状态引起的，这反过来又影响吸附的亲和力。这两种蛋白质都更强烈地结合到疏水表面，这种强烈的相互作用伴随着不太紧凑的构象。此外，可以看出，无论吸附剂表面的特性如何，两种蛋白质的双链体的旋转迁移率在吸附时都大大降低。(C)北京：科学出版社.
Interactions of recombinant human growth hormone and lysozyme with solid surfaces are studied using total internal reflection fluorescence (TIRF) and monitoring the protein's intrinsic tryptophan fluorescence. The intensity, spectra, quenching, and polarization of the fluorescence emitted by the adsorbed proteins are monitored and related to adsorption kinetics, protein conformation, and fluorophore rotational mobility. To study the influence of electrostatic and hydrophobic interactions on the adsorption process, three sorbent surfaces are used which differ in charge and hydrophobicity. The chemical surface groups are silanol, methyl, and quaternary amine. Results indicate that adsorption of hGH is dominated by hydrophobic interactions. Lysozyme adsoption is strongly affected by the ionic strength. This effect is probably caused by an ionic strength dependent conformational state in solution which, in turn, influences the affinity for adsorption. Both proteins are more strongly bound to hydrophobic surfaces and this strong interaction is accompanied by a less compact conformation. Furthermore, it was seen that regardless of the characteristics of the sorbent surface, the rotational mobility of both proteins' tryptophans is largely reduced upon adsorption. (C) 1997 Academic Press.