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

Giuseppe Santoro, Olivier Blacque, Fabio Zobi

Institute of Inorganic Chemistry, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.

Metallomics : integrated biometal science 2012 Mar

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The HEW lysozyme (Lys) and the fac-[Re(CO)(3)(H(2)O)(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. This journal is © The Royal Society of Chemistry 2012

Citation

Giuseppe Santoro, Olivier Blacque, Fabio Zobi. Post-protein binding metal-mediated coupling of an acridine orange-based fluorophore. Metallomics : integrated biometal science. 2012 Mar;4(3):253-9