Univ Lyon, Université Claude Bernard Lyon 1, CNRS, Institut Lumière Matière, UMR 5306, F-69622 Lyon, France.
Univ Lyon, Université Claude Bernard Lyon 1, CNRS, Institut Lumière Matière, UMR 5306, F-69622 Lyon, France.
Spectrochim Acta A Mol Biomol Spectrosc. 2018 Mar 15;193:283-288. doi: 10.1016/j.saa.2017.12.025. Epub 2017 Dec 9.
Protein-templated gold nanoclusters (AuNCs) are very attractive due to their unique fluorescence properties. A major problem however may arise due to protein structure changes upon the nucleation of an AuNC within the protein for any future use as in vivo probes, for instance. In this work, we propose a simple and reliable fluorescence based technique measuring the hydrodynamic size of protein-templated gold nanoclusters. This technique uses the relation between the time resolved fluorescence anisotropy decay and the hydrodynamic volume, through the rotational correlation time. We determine the molecular size of protein-directed AuNCs, with protein templates of increasing sizes, e.g. insulin, lysozyme, and bovine serum albumin (BSA). The comparison of sizes obtained by other techniques (e.g. dynamic light scattering and small-angle X-ray scattering) between bare and gold clusters containing proteins allows us to address the volume changes induced either by conformational changes (for BSA) or the formation of protein dimers (for insulin and lysozyme) during cluster formation and incorporation.
由于其独特的荧光性质,蛋白质模板金纳米团簇(AuNCs)非常有吸引力。然而,由于在蛋白质内形成 AuNC 时蛋白质结构的变化,可能会出现一个主要问题,例如,将来用作体内探针。在这项工作中,我们提出了一种简单可靠的基于荧光的技术,用于测量蛋白质模板金纳米团簇的流体力学尺寸。该技术通过旋转相关时间,利用荧光各向异性衰减与流体力学体积之间的关系。我们确定了具有不同大小蛋白质模板的蛋白质定向 AuNCs 的分子尺寸,例如胰岛素、溶菌酶和牛血清白蛋白(BSA)。通过其他技术(例如动态光散射和小角 X 射线散射)获得的 bare 和含金簇蛋白质之间的尺寸比较,使我们能够解决在簇形成和结合过程中由构象变化(对于 BSA)或蛋白质二聚体形成(对于胰岛素和溶菌酶)引起的体积变化。