Targeting the Surface of the Protein 14-3-3 by Ultrasmall (1.5 nm) Gold Nanoparticles Carrying the Specific Peptide CRaf.

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 (K D) 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.