Scanning Optical Spectroelectrochemistry: Applications in Protein Redox Potential Measurements

IF 6.1 Q1 CHEMISTRY, MULTIDISCIPLINARY
Prof. Paul V. Bernhardt
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引用次数: 1

Abstract

The accurate measurement of redox potentials of small molecules is a relatively straightforward task using electrochemical methods such as cyclic voltammetry. However, proteins, in most cases, are not amenable to the same approach due to slow heterogeneous electron transfer and the possibility of denaturing at the electrode surface. This necessitates the use of small molecular weight redox mediators to facilitate electron transfer. This leads to spectroelectrochemical techniques where the applied electrochemical potential is coupled to a spectroscopic signal of the protein. Traditionally this is done at different applied (fixed) potentials akin to an electrochemical titration, but the time required for electrochemical equilibrium to be established, and its consistent application, are major sources of experimental error. Here we have utilised a continuously scanning potential synchronised with time-resolved UV-vis spectroscopy to provide an automated approach that can be used to measure protein redox potentials accurately in an expedient manner. The test cases are the heme proteins cytochrome c and myoglobin. The scope and limitations of the method are discussed.

Abstract Image

扫描光谱电化学:在蛋白质氧化还原电位测量中的应用
使用循环伏安法等电化学方法精确测量小分子的氧化还原电位是一项相对简单的任务。然而,在大多数情况下,由于缓慢的非均相电子转移和电极表面变性的可能性,蛋白质不适合采用相同的方法。这就需要使用小分子量的氧化还原介质来促进电子转移。这导致了光谱电化学技术,其中应用的电化学电位与蛋白质的光谱信号耦合。传统上,这是在不同的应用(固定)电位下完成的,类似于电化学滴定,但建立电化学平衡所需的时间,以及它的一致性应用,是实验误差的主要来源。在这里,我们利用与时间分辨紫外可见光谱同步的连续扫描电位来提供一种自动化的方法,可以用一种方便的方式准确地测量蛋白质氧化还原电位。测试用例是血红蛋白、细胞色素c和肌红蛋白。讨论了该方法的适用范围和局限性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
7.30
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