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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
Author(s) -
Marcélis Lionel,
Kajouj Sofia,
Ghesquière Jonathan,
Fettweis Gregory,
Coupienne Isabelle,
Lartia Rémy,
Surin Mathieu,
Defrancq Eric,
Piette Jacques,
Moucheron Cécile,
KirschDe Mesmaeker Andrée
Publication year - 2016
Publication title -
european journal of inorganic chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.667
H-Index - 136
eISSN - 1099-0682
pISSN - 1434-1948
DOI - 10.1002/ejic.201600278
Subject(s) - chemistry , conjugate , hela , phototoxicity , confocal microscopy , ethidium bromide , peptide , flow cytometry , fluorescence microscope , confocal , cell penetrating peptide , biophysics , fluorescence , conjugated system , ruthenium , in vitro , dna , microbiology and biotechnology , biochemistry , organic chemistry , catalysis , mathematical analysis , physics , geometry , mathematics , quantum mechanics , biology , polymer
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 (ODN G ). Thus, the luminescence is quenched in the presence of ODN G , and gel electrophoresis experiments showed the appearance of products corresponding to the irreversible attachment of the conjugate to ODN G upon 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.