Targeting the Surface of the Protein 14-3-3 by Ultrasmall (1.5?nm) Gold Nanoparticles Carrying the Specific Peptide CRaf |
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| Authors: | Dr Tatjana Ruks Dr Kateryna Loza Dr Marc Heggen Prof?Dr Christian Ottmann Prof?Dr Peter Bayer Dr Christine Beuck Prof?Dr Matthias Epple |
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| Affiliation: | 1. Inorganic Chemistry and Center for Nanointegration Duisburg–Essen (CeNIDE), University of Duisburg-Essen, Universitätsstrasse 5–7, 45117 Essen, Germany;2. Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany;3. Laboratory of Chemical Biology Department of Biomedical Engineering and Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, P.O. Box 513, 5600MB Eindhoven, The Netherlands;4. Department of Structural and Medicinal Biochemistry, Centre for Medical Biotechnology (ZMB), University of Duisburg-Essen, 45117 Essen, Germany |
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| Abstract: | The surface of ultrasmall gold nanoparticles with an average diameter of 1.55 nm was conjugated with a 14-3-3 protein-binding peptide derived from CRaf. Each particle carries 18 CRaf peptides, leading to an overall stoichiometry of Au(115)Craf(18). The binding to the protein 14-3-3 was probed by isothermal titration calorimetry (ITC) and fluorescence polarization spectroscopy (FP). The dissociation constant (KD) was measured as 5.0 μM by ITC and 0.9 μM by FP, which was close to the affinity of dissolved CRaf to 14-3-3σ. In contrast to dissolved CRaf, which alone did not enter HeLa cells, CRAF-conjugated gold nanoparticles were well taken up by HeLa cells, opening the opportunity to target the protein inside a cell. |
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| Keywords: | Gold nanoparticles peptides proteins supramolecular chemistry |
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