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

Autor:	Jacques Piette, Eric Defrancq, Cécile Moucheron, Andrée Kirsch-De Mesmaeker, Mathieu Surin, Sofia Kajouj, Jonathan Ghesquière, Isabelle Coupienne, Rémy Lartia, Grégory Fettweis, Lionel Marcelis
Přispěvatelé:	Université Libre de Bruxelles, Université Libre de Bruxelles [Bruxelles] (ULB), Conception, synthèse et vectorisation de biomolécules. (CSVB), Institut Curie-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Département de Chimie Moléculaire - Ingéniérie et Intéractions BioMoléculaires (DCM - I2BM), Département de Chimie Moléculaire (DCM), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Laboratory for Chemistry of Novel Materials, Université de Mons (UMons), Université de Liège, Chimie Organique et Photochimie, Université libre de Bruxelles (ULB), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris], Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])
Rok vydání:	2016
Předmět:	Gel electrophoresis chemistry.chemical_classification medicine.diagnostic_test biology 010405 organic chemistry Stereochemistry Chemistry Peptide Cell sorting 010402 general chemistry biology.organism_classification 01 natural sciences 0104 chemical sciences Flow cytometry law.invention Inorganic Chemistry HeLa [CHIM.ANAL]Chemical Sciences/Analytical chemistry Confocal microscopy law medicine Biophysics Phototoxicity ComputingMilieux_MISCELLANEOUS Conjugate
Zdroj:	European Journal of Inorganic Chemistry European Journal of Inorganic Chemistry, Wiley-VCH Verlag, 2016, 2016 (18), pp.2902-2911. ⟨10.1002/ejic.201600278⟩
ISSN:	1099-0682 1434-1948
Popis:	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.
Databáze:	OpenAIRE
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