Characterization Of A Conformation-Restricted Amyloid Beta Peptide And Immunoreactivity Of Its Antibody In Human Ad Brain
ACS CHEMICAL NEUROSCIENCE(2021)
摘要
Characterization of amyloid beta (A beta) oligomers, the transition species present prior to the formation of A beta fibrils and that have cytotoxicity, has become one of the major topics in the investigations of Alzheimer's disease (AD) pathogenesis. However, studying pathophysiological properties of A beta oligomers is challenging due to the instability of these protein complexes in vitro. Here, we report that conformation-restricted A beta 42 with an intramolecular disulfide bond at positions 17 and 28 (SS-A beta 42) formed stable A beta oligomers in vitro. Thioflavin T binding assays, nondenaturing gel electrophoresis, and morphological analyses revealed that SS-A beta 42 maintained oligomeric structure, whereas wild-type A beta 42 and the highly aggregative A beta 42 mutant with E22P substitution (E22P-A beta 42) formed A beta fibrils. In agreement with these observations, SS-A beta 42 was more cytotoxic compared to the wild-type and E22P-A beta 42 in cell cultures. Furthermore, we developed a monoclonal antibody, designated TxCo-1, using the toxic conformation of SS-A beta 42 as immunogen. X-ray crystallography of the TxCo-1/SS-A beta 42 complex, enzyme immunoassay, and immunohistochemical studies confirmed the recognition site and specificity of TxCo-1 to SS-A beta 42. Immunohistochemistry with TxCo-1 antibody identified structures resembling senile plaques and vascular A beta in brain samples of AD subjects. However, TxCo-1 immunoreactivity did not colocalize extensively with A beta plaques identified with conventional A beta antibodies. Together, these findings indicate that A beta with a turn at positions 22 and 23, which is prone to form A beta oligomers, could show strong cytotoxicity and accumulated in brains of AD subjects. The SS-A beta 42 and TxCo-1 antibody should facilitate understanding of the pathological role of A beta with toxic conformation in AD.
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关键词
THP-1, A beta oligomer, protofibril, A beta fibril, A beta toxic conformer, Alzheimer's disease
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