Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Mar 3;588(5):701-4.
doi: 10.1016/j.febslet.2013.12.030. Epub 2014 Jan 17.

A simple method for the determination of reduction potentials in heme proteins

Igor Efimov  1 , Gary Parkin  1 , Elizabeth S Millett  1 , Jennifer Glenday  1 , Cheuk K Chan  1 , Holly Weedon  1 , Harpreet Randhawa  1 , Jaswir Basran  2 , Emma L Raven  3
Affiliations

A simple method for the determination of reduction potentials in heme proteins

Igor Efimov et al. FEBS Lett. .

Abstract

We describe a simple method for the determination of heme protein reduction potentials. We use the method to determine the reduction potentials for the PAS-A domains of the regulatory heme proteins human NPAS2 (Em=-115 mV ± 2 mV, pH 7.0) and human CLOCK (Em=-111 mV ± 2 mV, pH 7.0). We suggest that the method can be easily and routinely applied to the determination of reduction potentials across the family of heme proteins.

Keywords: Heme; Heme protein; Redox; Reduction potential.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
(A) Spectroscopic changes observed during the determination of the Fe3+/2+ reduction potential of the PAS-A domain of hNPAS2 using the dye Nile blue (25.0 °C, pH 7.0). Arrows indicate direction of absorbance changes. (B) The corresponding Nernst plot.
Fig. 2
Fig. 2
(A) Spectroscopic changes observed during the determination of the Fe3+/2+ reduction potential of the PAS-A domain of hCLOCK using the dye Nile blue (25.0 °C, pH 7.0). Arrows indicate direction of absorbance changes. (B) The corresponding Nernst plot.
See this image and copyright information in PMC

References

    1. Mauk A.G., Moore G.R. Control of metalloprotein redox potentials: what does site-directed mutagenesis of hemoproteins tell us? J. Biol. Inorg. Chem. 1997;2:119–125.
    1. Raven E.L., Mauk A.G. Chemical reactivity of the active site of myoglobin. Adv. Inorg. Chem. 2001;51:1–49.
    1. Battistuzzi G., Borsari M., Cowan J.A., Ranieri A., Sola M. Control of cytochrome C redox potential: axial ligation and protein environment effects. J. Am. Chem. Soc. 2002;124:5315–5324. - PubMed
    1. Pessanha M., Rothery E.L., Miles C.S., Reid G.A., Chapman S.K., Louro R.O., Turner D.L., Salgueiro C.A., Xavier A.V. Tuning of functional heme reduction potentials in Shewanella fumarate reductases. Biochim. Biophys. Acta. 2009;1787:113–120. - PubMed
    1. Bortolotti C.A., Amadei A., Aschi M., Borsari M., Corni S., Sola M., Daidone I. The reversible opening of water channels in cytochrome c modulates the heme iron reduction potential. J. Am. Chem. Soc. 2012;134:13670–13678. - PubMed

Publication types