A new method for quantitation of spin concentration by EPR spectroscopy: application to methemoglobin and metmyoglobin.
Abstract: A new method of EPR spectral analysis is developed to quantitate overlapping signals. The method requires double integration of a number of spectra containing the signals in different proportions and the subsequent solution of a system of linear equations. The result gives the double integral values of the individual lines, which can then be further used to find the concentrations of all the paramagnetic species present. There is no requirement to deconvolute the whole spectrum into its individual components. The method is employed to quantify different heme species in methemoglobin and metmyoglobin preparations. A significantly greater intensity of the high-spin signal in metmyoglobin, compared to methemoglobin at the same heme concentration, is shown to be due to larger amounts of low-spin forms in methemoglobin. Three low-spin types in methemoglobin and two in metmyoglobin are present in these samples. When their calculated concentrations are added to those of the high-spin forms, the results correspond to the total heme concentrations obtained by optical spectroscopy.
Copyright 2000 Academic Press.
Publication Date: 2000-01-29 PubMed ID: 10648142DOI: 10.1006/jmre.1999.1935Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research presents a novel method for measuring spin concentration in complex systems using EPR spectroscopy, highlighting its application in determining different forms of heme in methemoglobin and metmyoglobin.
Introduction to Research
- The research is aimed at developing a new approach for analyzing Electron Paramagnetic Resonance (EPR) spectral data to quantify overlapping signals. This method is then utilized to measure various heme species in methemoglobin and metmyoglobin preparations.
Methodology
- The newly developed method requires double integration of multiple spectra containing the signals in different proportions. From these integrated values, a system of linear equations is solved. This technique eliminates the need to deconvolute the complete spectrum into its individual elements.
- The solution to these linear equations provides the double integral values of the individual lines. These values can then be used to determine the concentrations of all the paramagnetic species present in the sample.
Findings
- Testing the method on methemoglobin and metmyoglobin, the researchers noticed a considerably higher intensity of the high-spin signal in metmyoglobin than in methemoglobin at the same heme concentration. This is attributed to the presence of more low-spin forms in methemoglobin.
- The study also identifies the presence of three low-spin types in methemoglobin and two in metmyoglobin.
- When the calculated concentrations of these spins were aggregated with those of the high-spin forms, the results aligned with the total heme concentrations obtained through optical spectroscopy, thus validating the new method.
Conclusion and Future Implications
- The study successfully developed and validated a novel approach for quantitation of spin concentration using EPR spectroscopy. The method provided a means of measuring components in complex systems without the need for complete spectral deconvolution, making it potentially useful in a variety of scientific applications.
Cite This Article
APA
Svistunenko DA, Sharpe MA, Nicholls P, Wilson MT, Cooper CE.
(2000).
A new method for quantitation of spin concentration by EPR spectroscopy: application to methemoglobin and metmyoglobin.
J Magn Reson, 142(2), 266-275.
https://doi.org/10.1006/jmre.1999.1935 Publication
Researcher Affiliations
- Department of Biological Sciences, Central Campus, University of Essex, Wivenhoe Park, Essex, CO4 3SQ, United Kingdom.
MeSH Terms
- Animals
- Electron Spin Resonance Spectroscopy / methods
- Heme / analysis
- Horses
- Humans
- Methemoglobin / chemistry
- Metmyoglobin / chemistry
Citations
This article has been cited 7 times.- Welbourn EM, Wilson MT, Yusof A, Metodiev MV, Cooper CE. The mechanism of formation, structure and physiological relevance of covalent hemoglobin attachment to the erythrocyte membrane. Free Radic Biol Med 2017 Feb;103:95-106.
- Otsuka M, Marks SA, Winnica DE, Amoscato AA, Pearce LL, Peterson J. Covalent modifications of hemoglobin by nitrite anion: formation kinetics and properties of nitrihemoglobin. Chem Res Toxicol 2010 Nov 15;23(11):1786-95.
- García-Rubio I, Braun M, Gromov I, Thöny-Meyer L, Schweiger A. Axial coordination of heme in ferric CcmE chaperone characterized by EPR spectroscopy. Biophys J 2007 Feb 15;92(4):1361-73.
- Dunne J, Caron A, Menu P, Alayash AI, Buehler PW, Wilson MT, Silaghi-Dumitrescu R, Faivre B, Cooper CE. Ascorbate removes key precursors to oxidative damage by cell-free haemoglobin in vitro and in vivo. Biochem J 2006 Nov 1;399(3):513-24.
- Svistunenko DA, Dunne J, Fryer M, Nicholls P, Reeder BJ, Wilson MT, Bigotti MG, Cutruzzolà F, Cooper CE. Comparative study of tyrosine radicals in hemoglobin and myoglobins treated with hydrogen peroxide. Biophys J 2002 Nov;83(5):2845-55.
- Svistunenko DA, Sharpe MA, Nicholls P, Blenkinsop C, Davies NA, Dunne J, Wilson MT, Cooper CE. The pH dependence of naturally occurring low-spin forms of methaemoglobin and metmyoglobin: an EPR study. Biochem J 2000 Nov 1;351 Pt 3(Pt 3):595-605.
- Reeder BJ, Svistunenko DA, Wilson MT. Hell's Gate Globin-I from Methylacidiphilum infernorum Displays a Unique Temperature-Independent pH Sensing Mechanism Utililized a Lipid-Induced Conformational Change. Int J Mol Sci 2024 Jun 20;25(12).
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
