Electrochemical titrations and reaction time courses monitored in situ by magnetic circular dichroism spectroscopy

被引:6
作者
Bradley, Justin M. [1 ]
Butt, Julea N. [1 ,2 ]
Cheesman, Myles R. [1 ]
机构
[1] Univ E Anglia, Ctr Mol & Struct Biochem, Sch Chem, Norwich NR4 7TJ, Norfolk, England
[2] Univ E Anglia, Ctr Mol & Struct Biochem, Sch Biol Sci, Norwich NR4 7TJ, Norfolk, England
基金
英国生物技术与生命科学研究理事会;
关键词
MCD; Magnetic circular dichroism; Electrochemistry; Metalloprotein; CYTOCHROME CD(1) NITRITE; NITROUS-OXIDE REDUCTASE; ELECTRONIC-STRUCTURE; PARACOCCUS-PANTOTROPHUS; THIOSPHAERA-PANTOTROPHA; GROUND-STATE; PSEUDOMONAS-STUTZERI; REDOX PROTEINS; COPPER SITES; C PEROXIDASE;
D O I
10.1016/j.ab.2011.07.030
中图分类号
Q5 [生物化学];
学科分类号
070307 [化学生物学];
摘要
Magnetic circular dichroism (MCD) spectra, at ultraviolet-visible or near-infrared wavelengths (185-2000 nm), contain the same transitions observed in conventional absorbance spectroscopy, but their bisignate nature and more stringent selection rules provide greatly enhanced resolution. Thus, they have proved to be invaluable in the study of many transition metal-containing proteins. For mainly technical reasons, MCD has been limited almost exclusively to the measurement of static samples. But the ability to employ the resolving power of MCD to follow changes at transition metal sites would be a potentially significant advance. We describe here the development of a cuvette holder that allows reagent injection and sample mixing within the 50-mm-diameter ambient temperature bore of an energized superconducting solenoid. This has allowed us, for the first time, to monitor time-resolved MCD resulting from in situ chemical manipulation of a metalloprotein sample. Furthermore, we report the parallel development of an electrochemical cell using a three-electrode configuration with physically separated working and counter electrodes, allowing true potentiometric titration to be performed within the bore of the MCD solenoid. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码:110 / 116
页数:7
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