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dc.contributor.authorDoyle, Colleen M.
dc.contributor.authorNaser, Dalia
dc.contributor.authorBauman, Heather A.
dc.contributor.authorRumfeldt, Jessica
dc.contributor.authorMeiering, Elizabeth M.
dc.date.accessioned2019-12-11 20:49:49 (GMT)
dc.date.available2019-12-11 20:49:49 (GMT)
dc.date.issued2019-08-15
dc.identifier.urihttps://doi.org/10.1016/j.ab.2019.03.007
dc.identifier.urihttp://hdl.handle.net/10012/15311
dc.descriptionThe final publication is available at Elsevier via https://doi.org/10.1016/j.ab.2019.03.007. © 2019. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.description.abstractBound metals are observed in a great many natural proteins, where they perform diverse roles in determining protein folding, stability and function. Due to the broad impact of bound metals on biophysical and biochemical properties of proteins, it is valuable to have accurate and facile methods for determining the metal content of proteins. Here we describe an optimized methodology using 4-(2-pyridylazo)resorcinol (PAR) to simultaneously quantify two metal ions in solution. The assay is demonstrated for quantification of Cu2+ and Zn2+ ions in human Cu, Zn superoxide dismutases (SOD1s); however, the method is general and can be applied to various combinations of metal ions. Advantages of the assay are that it is rapid and inexpensive, requires little sample and preparation, and has simple data analysis. We show that spectral decomposition software can accurately resolve the absorption bands of Cu2+ and Zn2+ with high accuracy and precision. Using the PAR assay, we determined that metal binding is altered in disease-associated mutants of SOD1, with comparable results to those determined by ICP-AES. In addition, we highlight key issues for using spectrophotometric chelators such as PAR for metal analysis of proteins.en
dc.description.sponsorshipThis work was supported by the Canadian Institutes of Health Research (CIHR) and National Sciences and Engineering Research Council of Canada (NSERC).en
dc.language.isoenen
dc.publisherElsevieren
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectmetalloproteinen
dc.subject4-(2-pyridylazo)resorcinolen
dc.subjectPARen
dc.subjectSOD1en
dc.subjectmetal quantitationen
dc.subjectassayen
dc.titleSpectrophotometric method for simultaneous measurement of zinc and copper in metalloproteins using 4-(2-pyridylazo)resorcinolen
dc.typeArticleen
dcterms.bibliographicCitationC. Doyle, D. Naser, H. Bauman, J. Rumfeldt, E. Meiering, Spectrophotometric method for simultaneous measurement of zinc and copper in metalloproteins using 4-(2- pyridylazo)resorcinol, Analytical Biochemistry (2019), doi: https://doi.org/10.1016/j.ab.2019.03.007.en
uws.contributor.affiliation1Faculty of Scienceen
uws.contributor.affiliation2Chemistryen
uws.typeOfResourceTexten
uws.peerReviewStatusRevieweden
uws.scholarLevelFacultyen
uws.scholarLevelPost-Doctorateen
uws.scholarLevelGraduateen


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