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  • Title: Conjugation of Au11 cluster with Cys-rich peptides containing the alpha-domain of metallothionein.
    Author: Ariyasu S, Onoda A, Sakamoto R, Yamamura T.
    Journal: Dalton Trans; 2009 May 21; (19):3742-7. PubMed ID: 19417939.
    Abstract:
    The stoichiometrically controlled complexation of abiological inorganic clusters with proteins is a challenging task that enables us to combine the functions of inorganic clusters with those of biological systems on the basis of chemical characterisation. In this study, we synthesised a 1:1 conjugate between Au11(PPh3)8Cl3 (Au11) and CGMTIIalpha, where Au11 is the smallest cluster, which is situated on the boundary of molecules and nanoparticles, having discrete electronic states and exhibiting Coulomb blockade; on the other hand, CGMTIIalpha is a twelve-cysteine (Cys)-containing peptide consisting of the alpha-domain of rat liver metallothionein II (MTIIalpha)--a heavy metal-binding protein--and an addendum, cysteine-glycine (CG). The Cys residue of the N-terminus is used as a site for elongation by the native chemical ligation (NCL) method. Titration experiments monitored by circular dichroism spectroscopy indicate that CGMTIIalpha reacts with Au11 in a ratio of 1:1. The product exhibits a negative Cotton effect at 222 nm, indicating the formation of a short a-helix. The composition of this hybrid module is quantified as Au11(PPh3)4(S-Cys)6 by transmission electron microscopy (core size observation), energy-dispersive X-ray spectroscopy (elemental analysis), Ellman test (uncoordinated Cys residues), and high-performance liquid chromatography (quantification of eliminated PPh3). These results, as well as those obtained from the simulations of Au 4f X-ray photoelectron spectra, suggest that the Au cluster of this product is Au11(PPh3)4(S-Cys)6. We also prepared F3F3-CGMTIIalpha, where F3F3 is a zinc finger peptide with two domains, in order to demonstrate the applicability of the NCL method to the elongation of CGMTIIalpha. F3F3-CGMTIIalpha also reacts with Au11 to yield a 1:1 conjugate without interfering in the folding of the zinc finger.
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