An intramolecular O-N migration reaction on gold surfaces: toward the preparation of well-defined amyloid surfaces.
Amyloids are a family of self-aggregating proteins implicated in various central nervous system disorders, including Alzheimer's disease (AD). It is thought that prefibrillar soluble forms of amyloid peptides, including oligomers, may be the main pathogenic factor in AD. Herein we describe the fabrication of well-defined, functionalized, monomeric beta-amyloid peptide surfaces for studying protein-protein interactions. We first prepared a nonaggregating analogue of the beta-amyloid peptide and then attached it to a gold surface covered with a self-assembled monolayer (SAM) of alkanethiols. After attachment, the native form of the beta-amyloid peptide (Abeta40) was obtained by surface-level intramolecular O-N migration. The surface was characterized by atomic force microscopy (AFM) and self-assembled monolayer for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (SAMDI-TOF MS). The interaction between the surface-bound Abeta40 and monoclonal anti-Abeta40 antibody was tracked by AFM and chemiluminescence, which revealed that the Abeta40 was attached mainly in its monomeric form and that the protein-protein complex was assembled on the surface. Last, we used a proteomics approach to demonstrate the specificity of the Abeta40-functionalized surface in surface-binding experiments employing serum amyloid P (SAP) and bovine serum albumin (BSA).
Boussert
Stephanie
S
Proteomics Platform, Barcelona Science Park, Baldiri Reixac 10, E-08028 Barcelona, Spain.
Diez-Perez
Ismael
I
Kogan
Marcelo J
MJ
de Oliveira
Eliandre
E
Giralt
Ernest
E
D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D002417:Cattle; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000737:chemistry*; D010100:Oxygen / Q000737:chemistry*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D000683:Serum Amyloid P-Component / Q000737:chemistry / Q000378:metabolism; D013499:Surface Properties
D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; D000683:Serum Amyloid P-Component; C078592:amyloid beta-protein (1-40); D012710:Serum Albumin, Bovine; D006046:Gold; D009584:Nitrogen; D010100:Oxygen