In vitro formation of amyloid fibrils from two synthetic peptides of different lengths homologous to alzheimer's disease β-protein

Two synthetic peptides corresponding to the reported 28-residue sequence of Alzheimer's Disease β-protein (SP28) and to residues 12-28 (SP17) were used to form fibrils in vitro. Synthetic fibrils bound Congo Red and closely resembled amyloid fibrils isolated from leptomeninges and senile plaques of Alzheimer's brain by electron microscopy. A polyclonal antiserum to SP28 specifically decorated both synthetic and native amyloid by colloidal gold immunoelectron microscopy. Amyloid fibrils isolated from tissue were insoluble on SDS-Polyacrylamide gels, and tended to aggregate while synthetic amyloid fibrils were completely solubilized, releasing only monomers of SP28 and SP17. Anti-SP28 immunostained cerebrovascular and plaque core amyloid, but not neurofibrillary tangles, in tissue section. Western blot analysis showed that anti-SP28 reacted with a 4 kDa band released from amyloid core enriched preparations and leptomeninges. By contrast, a 16 kDa band corresponding to the tetramer of β-protein was not recognized. These data suggest that as little as a 17 residue sequence of β-protein may be required to form fibrils and that the complete sequence of the 4 kDa β-protein may be important in determining insolubility and the formation of intermediate size polymers.