A cell-penetrating artificial metalloenzyme regulates a gene switch in a designer mammalian cell
JournalArticle (Originalarbeit in einer wissenschaftlichen Zeitschrift)
ID 4496053
Author(s) Okamoto, Yasunori; Kojima, Ryosuke; Schwizer, Fabian; Bartolami, Eline; Heinisch, Tillmann; Matile, Stefan; Fussenegger, Martin; Ward, Thomas R.
Author(s) at UniBasel Ward, Thomas R.
Okamoto, Yasunori
Schwizer, Fabian
Heinisch, Tillmann
Fussenegger, Martin
Year 2018
Title A cell-penetrating artificial metalloenzyme regulates a gene switch in a designer mammalian cell
Journal Nature Communications
Volume 9
Pages / Article-Number 1943
Abstract Complementing enzymes in their native environment with either homogeneous or heterogeneous catalysts is challenging due to the sea of functionalities present within a cell. To supplement these efforts, artificial metalloenzymes are drawing attention as they combine attractive features of both homogeneous catalysts and enzymes. Herein we show that such hybrid catalysts consisting of a metal cofactor, a cell-penetrating module, and a protein scaffold are taken up into HEK-293T cells where they catalyze the uncaging of a hormone. This bioorthogonal reaction causes the upregulation of a gene circuit, which in turn leads to the expression of a nanoluc-luciferase. Relying on the biotin-streptavidin technology, variation of the biotinylated ruthenium complex: the biotinylated cell-penetrating poly(disulfide) ratio can be combined with point mutations on streptavidin to optimize the catalytic uncaging of an allyl-carbamate-protected thyroid hormone triiodothyronine. These results demonstrate that artificial metalloenzymes offer highly modular tools to perform bioorthogonal catalysis in live HEK cells.
Publisher Nature Research
ISSN/ISBN 2041-1723
edoc-URL https://edoc.unibas.ch/70534/
Full Text on edoc Available
Digital Object Identifier DOI 10.1038/s41467-018-04440-0
ISI-Number WOS:000432279600004
Document type (ISI) Article

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