Copper(I) and copper(II) binding to b-amyloid 16 (Ab16) studied by electrospray ionization mass spectrometryw

Copper-b-amyloid 16 (Ab16) complexes were investigated by electrospray ionization mass spectrometry (ESI-MS). Copper(I) and (II) complexes were formed on-line in a microchip electrospray emitter by using a sacrificial copper electrode as the anode in positive ionization mode. In the presence of ascorbic acid in the peptide solution, the amount of Cu(I)-Ab16 generated electrochemically was even higher. A kinetic model is proposed to account for the generation of copper complexes. The structure of Cu(I)-Ab16 was investigated by tandem mass spectrometry (MS/MS), and the binding site of Cu(I) to Ab16 was identified at the His13, His14 residues. Cu(II)-Ab16 was also investigated by MS/MS and, based on the unusual observations of a-ions, the two binding residues of His13 and His14 of Ab16 to Cu(II) were also confirmed. This approach provides direct information on Cu(I)-Ab16 complexes generated in solution from metallic copper and gives evidence that both His13 and His14 are involved in the coordination of both Cu(I)and Cu(II)-Ab16 complexes.