Highly DNA-Photoreactive Ruthenium 1,4,5,8-Tetraazaphenanthrene Complex Conjugated to the TAT Peptide: Efficient Vectorization inside HeLa Cells without Phototoxicity – The Importance of Cellular Distribution
[en] The photoreactive [Ru(TAP)2(phen)]2+(TAP = 1,4,5,8-tetraazaphenanthrene; phen = 1,10-phenanthroline) complex tethered to the cell-penetrating peptide (CPP) TAT was studied in vitro and in cellulo. The tethering of the complex does not affect its behavior under blue-light irradiation in the presence of guanine-containing oligodeoxyribonucleotides (ODNG). Thus, the luminescence is quenched in the presence of ODNG, and gel electrophoresis experiments showed the appearance of products corresponding to the irreversible attachment of the conjugate to ODNGupon illumination. The cellular uptake of the conjugate was examined by flow cytometry, inductively coupled plasma mass spectrometry (ICP-MS), and confocal imaging microscopy. These experiments showed that the [Ru(TAP)2(phen-TAT)] conjugate is readily taken up by HeLa cells and, despite these favorable factors, the cellular survival was 100 %, as measured by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. A possible origin of the inactivity of [Ru(TAP)2(phen-TAT)] under irradiation is proposed on the basis of the fluorescence-activated cell sorting (FACS), ICP-MS, and confocal microscopy results.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Marcélis, Lionel ; Organic Chemistry and Photochemistry, Université libre de Bruxelles (U.L.B.), Bruxelles, Belgium
Kajouj, Sofia ; Organic Chemistry and Photochemistry, Université libre de Bruxelles (U.L.B.), Bruxelles, Belgium
Ghesquière, Jonathan; Organic Chemistry and Photochemistry, Université libre de Bruxelles (U.L.B.), Bruxelles, Belgium
Fettweis, Grégory ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Virologie - Immunologie
Coupienne, Isabelle ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Virologie - Immunologie
Lartia, Rémy; Département de Chimie Moléculaire, UMR CNRS, Université Grenoble Alpes, Grenoble, France
Surin, Mathieu; Laboratory for Chemistry of Novel Materials, UMR CNRS, University of Mons - UMons, Mons, Belgium
Defrancq, Eric ; Département de Chimie Moléculaire, UMR CNRS, Université Grenoble Alpes, Grenoble, France
Piette, Jacques ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Virologie et immunologie
Moucheron, Cécile ; Organic Chemistry and Photochemistry, Université libre de Bruxelles (U.L.B.), Bruxelles, Belgium
Kirsch-De Mesmaeker, Andrée ; Organic Chemistry and Photochemistry, Université libre de Bruxelles (U.L.B.), Bruxelles, Belgium
Language :
English
Title :
Highly DNA-Photoreactive Ruthenium 1,4,5,8-Tetraazaphenanthrene Complex Conjugated to the TAT Peptide: Efficient Vectorization inside HeLa Cells without Phototoxicity – The Importance of Cellular Distribution
The authors gratefully thank N. Matielli and J. De Jong (G-Time Lab at the U. L. B.) for the ICP-MS measurements. We also acknowledge gratefully Professor Guillermo Orellana for the measurements of singlet-oxygen quantum yields realized at the Universidad Complutense de Madrid. This research was supported by the Interuniversity Attraction Poles Programme (P7/05 and P07/32) of the Belgian Science Policy Office and by the “Fonds National pour la Recherche Scientifique” (FNRS) under grant numbers 2.4615.11 and 2.4530.12. The Nanobio-ICMG platform (FR 2607) in Grenoble is acknowledged for providing facilities for the synthesis and purification of conjugates. L. M. and J. G. are grateful to the Fonds de la Recherche Scientifique – FNRS (F.R.S.-FNRS) for a PhD grant. S. K. thanks the Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture (F.R.I.A.) for a PhD grant. M. S. is Research Associate of the F.R.S.-FNRS. G. F. was supported by a grant from the Télévie (F.R.S.-FNRS).
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