Please use this identifier to cite or link to this item:
|Title:||Monitoring Changes in the Redox State of Myoglobin in Cardiomyocytes by Raman Spectroscopy Enables the Protective Effect of NO Donors to Be Evaluated|
Kapetanaki, Sofia M.
Storey, Nina M.
|Publisher:||American Chemical Society|
|Citation:||Analytical Chemistry, 2015, 87 (20), pp 10605–10612|
|Abstract:||Raman microspectroscopy has been used to monitor changes in the redox and ligand-coordination states of the heme complex in myoglobin during the pre-conditioning of ex vivo cardiomyocytes with pharmacological drugs that release nitric oxide (NO). These chemical agents are known to confer protection on heart tissue against ischemia-reperfusion injury. Subsequent changes in the redox and ligand-coordination states during experimental simulations of ischemia and reperfusion have also been monitored. We found that these measurements, in real time, could be used to evaluate the pre-conditioning treatment of cardiomyocytes, and predict the likelihood of cell survival following a potentially-lethal period of ischemia. Evaluation of the pre-conditioning treatment was done at the single-cell level. The binding of NO to myoglobin, giving a 6-coordinate ferrous-heme complex, was inferred from the measured Raman bands of a cardiomyocyte by comparison to pure solution of the protein in the presence of NO. A key change in the Raman spectrum was observed after perfusion of the NO-donor was completed, where if the pre-conditioning treatment was successful then the bands corresponding to the nitrosyl complex were replaced by bands corresponding to metmyoglobin, Mb[SUPERSCRIPT: III]. An observation of Mb[SUPERSCRIPT: III] bands in the Raman spectrum was made for all the cardiomyocytes that recovered contractile function, whilst the absence of Mb[SUPERSCRIPT: III] bands always indicated that the cardiomyocyte would be unable to recover contractile function, following the simulated conditions of ischemia and reperfusion in these experiments.|
|Rights:||Copyright © 2015 American Chemical Society. This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see DOI: 10.1021/acs.analchem.5b03103|
|Description:||The file associated with this record is under a 12-month embargo from publication in accordance with the publisher's self-archiving policy, available at http://pubs.acs.org/page/4authors/jpa/index.html|
|Appears in Collections:||Published Articles, Dept. of Chemistry|
Files in This Item:
|Manuscript 210912.docx||Post-review (final submitted)||2.24 MB||Unknown||View/Open|
|Manuscript 210912.pdf||Post-review (final submitted)||4.03 MB||Adobe PDF||View/Open|
Items in LRA are protected by copyright, with all rights reserved, unless otherwise indicated.