Copper(II) interaction with peptide fragments of histidine–proline-rich glycoprotein: Speciation, stability and binding details

Abstract

GHHPH is the peptide repeat present in histidine–proline rich glycoprotein (HPRG), a plasma glycoprotein involved in angiogenesis process. The copper(II) ions interaction with mono (Ac-GHHPHG-NH2) and its bis-repeat (Ac-GHHPHGHHPHG-NH2) was investigated by means of potentiometric and spectroscopic techniques. To single out the copper(II) coordination environments of different species formed with Ac-GHHPHG-NH2, three single point mutated peptides were also synthesized and their ability to coordinate Cu2+ investigated. Ac-GHHPHG-NH2 binds Cu2+ by the imidazole side chain and the amide nitrogen deprotonation that takes place towards the N-terminus. The bis-repeat is able to bind Cu2+ more efficiently than Ac-GHHPHG-NH2. This difference is not only due to the number of His residues in the sequence but also to the different binding sites. In fact, the comparison of the potentiometric and spectroscopic data of the copper(II) complexes with a bis-repeatPeg construct Ac-(GHHPHG)-Peg-(GHHPHG)-NH2 and those of the metal complexes with Ac-HGHH-NH2, indicates that the central HGHH amino acid sequence is the main copper(II) binding site.