Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, AB, T6G 2G2, Canada.
Anal Biochem. 2021 Sep 15;629:114269. doi: 10.1016/j.ab.2021.114269. Epub 2021 Jun 3.
The near universal availability of UV-Visible spectrophotometers makes this instrument a highly exploited tool for the inexpensive, rapid examination of iron-sulfur clusters. Yet, the analysis of iron-sulfur cluster reconstitution experiments by UV-Vis spectroscopy is notoriously difficult due to the presence of broad, ill-defined peaks. Other types of spectroscopies, such as electron paramagnetic resonance spectroscopy and Mössbauer spectroscopy, are superior in characterizing the type of cluster present and their associated electronic transitions but require expensive, less readily available equipment. Here, we describe a tool that utilizes the accessible and convenient platform of Microsoft Excel to allow for the semi-quantitative analysis of iron-sulfur clusters by UV-Vis spectroscopy. This tool, which we call Fit-FeS, could potentially be used to additionally decompose spectra of solutions containing chromophores other than iron-sulfur clusters.
近紫外线可见分光光度计的广泛应用使该仪器成为一种廉价、快速检测铁硫簇的常用工具。然而,由于存在宽而无明确界限的峰,因此通过紫外可见光谱法分析铁硫簇的重组实验非常困难。其他类型的光谱学,如电子顺磁共振波谱和穆斯堡尔谱学,在表征存在的簇类型及其相关的电子跃迁方面更为优越,但需要昂贵、不太容易获得的设备。在这里,我们描述了一种工具,它利用微软 Excel 这个易于访问和方便的平台,允许通过紫外可见光谱对铁硫簇进行半定量分析。我们称之为 Fit-FeS 的这个工具,可能还可以用于分解除铁硫簇以外的发色团溶液的光谱。