Spectroscopic and electronic structure studies of copper(II) binding to His111 in the human prion protein fragment 106-115: evaluating the role of protons and methionine residues.
The prion protein (PrP(C)) is implicated in the spongiform encephalopathies in mammals, and it is known to bind Cu(II) at the N-terminal region. The region around His111 has been proposed to be key for the conversion of normal PrP(C) to its infectious isoform PrP(Sc). The principal aim of this study is to understand the role of protons and methionine residues 109 and 112 in the coordination of Cu(II) to the peptide fragment 106-115 of human PrP, using different spectroscopic techniques (UV-vis absorption, circular dichroism, and electron paramagnetic resonance) in combination with detailed electronic structure calculations. Our study has identified a proton equilibrium with a pK(a) of 7.5 associated with the Cu(II)-PrP(106-115) complex, which is ascribed to the deprotonation of the Met109 amide group, and it converts the site from a 3NO to a 4N equatorial coordination mode. These findings have important implications as they imply that the coordination environment of this Cu binding site at physiological pH is a mixture of two species. This study also establishes that Met109 and Met112 do not participate as equatorial ligands for Cu, and that Met112 is not an essential ligand, while Met109 plays a more important role as a weak axial ligand, particularly for the 3NO coordination mode. A role for Met109 as a highly conserved residue that is important to regulate the protonation state and redox activity of this Cu binding site, which in turn would be important for the aggregation and amyloidogenic properties of the protein, is proposed.
Departamento de Química, Centro de Investigación y de Estudios Avanzados (Cinvestav), 07360, DF, Mexico.
Lina
L
Rivillas-Acevedo
Rafael
R
Grande-Aztatzi
Italia
I
Lomelí
Javier E
JE
García
Erika
E
Barrios
Sarai
S
Teloxa
Alberto
A
Vela
Liliana
L
Quintanar
D002942:Circular Dichroism; D003300:Copper / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D006639:Histidine / Q000378:metabolism*; D006801:Humans; D008715:Methionine / Q000378:metabolism*; D011328:Prions / Q000737:chemistry / Q000378:metabolism*; D011522:Protons; D013056:Spectrophotometry, Ultraviolet
D011328:Prions; D011522:Protons; D006639:Histidine; D003300:Copper; D008715:Methionine