Nickel(II) binding to Cap43 protein fragments

Abstract

Cap43 protein has been tested for metal binding domains. The protein, specifically induced by nickel compounds in cultured human cells, had a new mono-histidinic motif consisting of 10 amino acids repeated three times in the C-terminus. The 20-Ac-TRSRSHTSEG–TRSRSHTSEG (Thr 341–Arg–Ser–Arg–Ser–His 346–Thr–Ser–Glu–Gly–Thr–Arg–Ser–Arg–Ser–His 356–Thr–Ser–Glu–Gly 360 – peptide 1) and the 30–Ac-TRSRSHTSEG–TRSRSHTSEG–TRSRSHTSEG (Thr 341–Arg–Ser–Arg–Ser–His 346–Thr–Ser–Glu–Gly–Thr–Arg–Ser–Arg–Ser–His 356–Thr–Ser–Glu–Gly–Thr–Arg–Ser–Arg–Ser–His 366–Thr–Ser–Glu–Gly 370 – peptide 2) amino acids sequence has been analyzed as a site for Ni(II) binding. A combined pH-metric and spectroscopic (UV–visible, CD, NMR) studies of Ni(II) binding to both fragments were performed. The 20-amino acid peptide can bind one and two metal ions while the 30-amino acid fragment one, two and three metal ions. At physiological pH, depending on the metal to ligand molar ratio, peptide 1 forms the Ni 2L species while peptide 2 the NiL, Ni 2L and Ni 3L complexes where each metal ion is coordinated to the imidazole nitrogen atom of the histidine residue of the 10-amino acid fragment. Octahedral complexes at pH 8–9 and planar 4N complexes with (N Im, 3N −) bonding mode at pH above 9, are formed. This work supports the existence of an interesting binding site at the COOH-terminal domain of the Cap43 protein.