Methionine Does Not Reduce Cu(II)-b-Amyloid! —Rectification of the Roles of Methionine-35 and Reducing Agents in Metal-Centered Oxidation Chemistry of Cu(II)-β-Amyloid

Giordano F. Z. da Silva,a# Vasiliky Lykourinou,a Alexander Angerhofer,b* and Li-June Minga*

a  Department of Chemistry and Institute for Biomolecular Science, University of South Florida, Tampa, Florida 33620-5250. 

b Department of Chemistry, University of Florida, Gainesville, Florida 32611



            The potential risk of metal-centered oxidative catalysis has been overlooked in the research of the copper complexes of the Alzheimer’s disease-related β-amyloid (Aβ) peptides.  Cu2+ complexes of Aβ1–40 and its 1–16 and 1–20 fragments have recently been shown to exhibit significant metal-centered oxidative activities toward several catecholamine neurotransmitters with and without H2O2 around neutral pH [G.F.Z da Silva, L.-J. Ming, Angew. Chem. Int. Ed. 46 (2007) 3337].  The results further support the metallo-Aβ-associated oxidative stress theory often considered to be connected to the neuropathology of the disease.  The metal-centered oxidative catalysis of CuAβ1-16/20 challenges the long-standing proposed redox role of Met35 in Aβ because Aβ1–16/20 do not contain a Met.  External Met has been determined by kinetic, optical, and electron paramagnetic resonance methods to bind directly to the Cu2+ center of CuAβ1–40 and CuAβ1–20 with Kd = 2.8 mM and 11.3 mM, respectively, which reflects less accessibility of the metal center in the full-length CuAβ1–40.  However, Met does not serve as a reducing agent for the Cu(II) which thus must amplify the observed oxidative catalysis of CuAβ1–20 through a non-redox mechanism.  Conversely, the CuAβ-catalyzed oxidation reaction of dopamine is inhibited by bio-available reducing agents such as ascorbate (competitive Kic = 66 μM) and glutathione (non-competitive, Kinc = 53 μM).  These data indicate that the oxidation chemistry of metallo-Ab is not initiated by Met35.  The results yield further molecular and mechanistic insights into the roles of metallo-Aβ in this disease.