Structural alterations in the peptide backbone of β-amyloid core protein may account for its deposition and stability in Alzheimer's disease

The structure of β-amyloid (βA) from Alzheimer disease brains was examined to determine if post-translational modifications might be linked to the abnormal deposition of this peptide in the diseased tissue. The βA peptides were isolated from the compact amyloid cores of neuritic plaques and separated from minor glycoprotein components by size-exclusion high-pressure liquid chromatography (HPLC). This parenchymal βA has a maximal length of 42 residues, but shorter forms with "ragged" NH₂ termini are also present. Tryptic peptide analysis revealed heterogeneity in the βA^1-5 and βA^6-16 peptides, each of which eluted as four peaks on reverse phase HPLC. Amino acid composition and sequence analyses, mass spectrometry, enzymatic methylation, and stereoisomer determinations revealed that these multiple peptide forms resulted from structural rearrangements of the aspartyl residues at βA positions 1 and 7. The L-isoaspartyl form predominates at each of these positions, whereas the D-isoaspartyl, L-aspartyl, and D-aspartyl forms are present in lesser amounts. βA purified from the leptomeningeal microvasculature contains the same structural alterations as parenchymal βA, but is 2 residues shorter at its COOH terminus. Using two different purification protocols, and using a synthetic βA^1-42 peptide as a control, we show that these modifications arose endogenously and were not caused by the experimental manipulations. The abundance of structurally altered aspartyl residues may profoundly affect the conformation of the βA protein within plaque cores and thus significantly impact normal catabolic processes designed to limit its deposition. These alterations may therefore contribute to the production and stability of β-amyloid deposits in Alzheimer brain tissue.