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Post-protein-binding metal-mediated coupling of an acridine orange-based fluorophore


Santoro, G; Blacque, O; Zobi, F (2012). Post-protein-binding metal-mediated coupling of an acridine orange-based fluorophore. Metallomics, 4(3):253-259.

Abstract

The HEW lysozyme (Lys) and the fac-[Re(CO)3(H2O)3]+ complex (1) are used as a simple model system for the description of a new approach to the labelling polypeptides with fluorescent tags. The strategy takes advantage of the reaction of an acridine orange-based fluorophore (AO) with the non-native metal fragment 1 hybridized on the enzyme. A synthetic methodology for the quantitative metallation of the protein is first described and it is then shown that the exogenous metal complex can be exploited for the coupling of the fluorescent probe. All Lys-derived species were characterized by various spectroscopic techniques. It is shown that the approach does not significantly alter the activity of the final fluorescent metallo-protein conjugate (Lys2). The accumulation of Lys2 on Micrococcus lysodeikticus bacteria was observed via confocal laser scanning microscopy.

Abstract

The HEW lysozyme (Lys) and the fac-[Re(CO)3(H2O)3]+ complex (1) are used as a simple model system for the description of a new approach to the labelling polypeptides with fluorescent tags. The strategy takes advantage of the reaction of an acridine orange-based fluorophore (AO) with the non-native metal fragment 1 hybridized on the enzyme. A synthetic methodology for the quantitative metallation of the protein is first described and it is then shown that the exogenous metal complex can be exploited for the coupling of the fluorescent probe. All Lys-derived species were characterized by various spectroscopic techniques. It is shown that the approach does not significantly alter the activity of the final fluorescent metallo-protein conjugate (Lys2). The accumulation of Lys2 on Micrococcus lysodeikticus bacteria was observed via confocal laser scanning microscopy.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Chemistry
Dewey Decimal Classification:540 Chemistry
Language:English
Date:2012
Deposited On:04 Mar 2013 14:31
Last Modified:05 Apr 2016 16:40
Publisher:RSC Publishing
ISSN:1756-5901
Additional Information:Persons who receive the PDF must not make it further available or distribute it.
Publisher DOI:https://doi.org/10.1039/C2MT00175F

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