Covalent Attachment of Cyclic TAT Peptides to GFP Results in Protein Delivery into Live Cells with Immediate Bioavailability |
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| Authors: | Dr Francesco Natale Nina Bohlke Prof Nediljko Budisa Prof M Cristina Cardoso Prof Christian P R Hackenberger |
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| Institution: | 1. Technische Universit?t Darmstadt, Fachbereich Biologie, Schnittspahnstrasse 10, 64287 Darmstadt (Germany);2. Technische Universit?t Berlin, Institut für Chemie, Müller‐Breslau‐Str. 10, 10623 Berlin (Germany);3. Technische Universit?t Darmstadt, Fachbereich Biologie, Schnittspahnstrasse 10, 64287 Darmstadt (Germany)These authors contributed equally.;4. Leibniz‐Institut für Molekulare Pharmakologie (FMP), Robert‐R?ssle‐Strasse 10, 13125 Berlin (Germany);5. Humboldt Universit?t zu Berlin, Department Chemie, Brook‐Taylor‐Strasse 2, 12489 Berlin (Germany)These authors contributed equally. |
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| Abstract: | The delivery of free molecules into the cytoplasm and nucleus by using arginine‐rich cell‐penetrating peptides (CPPs) has been limited to small cargoes, while large cargoes such as proteins are taken up and trapped in endocytic vesicles. Based on recent work, in which we showed that the transduction efficiency of arginine‐rich CPPs can be greatly enhanced by cyclization, the aim was to use cyclic CPPs to transport full‐length proteins, in this study green fluorescent protein (GFP), into the cytosol of living cells. Cyclic and linear CPP–GFP conjugates were obtained by using azido‐functionalized CPPs and an alkyne‐functionalized GFP. Our findings reveal that the cyclic‐CPP–GFP conjugates are internalized into live cells with immediate bioavailability in the cytosol and the nucleus, whereas linear CPP analogues do not confer GFP transduction. This technology expands the application of cyclic CPPs to the efficient transport of functional full‐length proteins into live cells. |
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| Keywords: | cell‐penetrating peptides click chemistry cyclic peptides live‐cell microscopy protein delivery |
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