Thyparambil Aby A, Wei Yang, Latour Robert A
Department of Bioengineering, Clemson University, 501 Rhodes Engineering Research Center, Clemson, South Carolina 29634.
Biointerphases. 2015 Mar 30;10(1):019002. doi: 10.1116/1.4906485.
Protein adsorption on material surfaces is a common phenomenon that is of critical importance in many biotechnological applications. The structure and function of adsorbed proteins are tightly interrelated and play a key role in the communication and interaction of the adsorbed proteins with the surrounding environment. Because the bioactive state of a protein on a surface is a function of the orientation, conformation, and accessibility of its bioactive site(s), the isolated determination of just one or two of these factors will typically not be sufficient to understand the structure-function relationships of the adsorbed layer. Rather a combination of methods is needed to address each of these factors in a synergistic manner to provide a complementary dataset to characterize and understand the bioactive state of adsorbed protein. Over the past several years, the authors have focused on the development of such a set of complementary methods to address this need. These methods include adsorbed-state circular dichroism spectropolarimetry to determine adsorption-induced changes in protein secondary structure, amino-acid labeling/mass spectrometry to assess adsorbed protein orientation and tertiary structure by monitoring adsorption-induced changes in residue solvent accessibility, and bioactivity assays to assess adsorption-induced changes in protein bioactivity. In this paper, the authors describe the methods that they have developed and/or adapted for each of these assays. The authors then provide an example of their application to characterize how adsorption-induced changes in protein structure influence the enzymatic activity of hen egg-white lysozyme on fused silica glass, high density polyethylene, and poly(methyl-methacrylate) as a set of model systems.
蛋白质在材料表面的吸附是一种常见现象，在许多生物技术应用中至关重要。吸附蛋白质的结构和功能紧密相关，在吸附蛋白质与周围环境的通讯和相互作用中起关键作用。由于蛋白质在表面的生物活性状态是其生物活性位点的取向、构象和可及性的函数，仅孤立地测定这些因素中的一两个通常不足以理解吸附层的结构-功能关系。相反，需要结合多种方法以协同方式处理这些因素中的每一个，以提供补充数据集来表征和理解吸附蛋白质的生物活性状态。在过去几年中，作者专注于开发这样一套补充方法以满足这一需求。这些方法包括吸附态圆二色光谱偏振法，用于确定蛋白质二级结构的吸附诱导变化；氨基酸标记/质谱法，通过监测吸附诱导的残基溶剂可及性变化来评估吸附蛋白质的取向和三级结构；以及生物活性测定法，用于评估吸附诱导的蛋白质生物活性变化。在本文中，作者描述了他们为每种测定开发和/或改编的方法。然后，作者提供了一个应用示例，以表征蛋白质结构的吸附诱导变化如何影响作为一组模型系统的熔融石英玻璃、高密度聚乙烯和聚（甲基丙烯酸甲酯）上的鸡蛋清溶菌酶的酶活性。
