title: Protein L-isoaspartyl O-methyltransferase inhibits amyloid beta fibrillogenesis in vitro.
abstract: Fibrillar aggregates of beta-amyloid peptide (Abeta) are major constituents of the senile plaques found in the brains of patients suffering from Alzheimer's disease (AD). Previous studies have shown that spontaneous isomerization or racemization of aspartyl residues in Abeta peptides leads to conformational changes in the secondary structure and increased aggregative ability of the peptides. Protein L-isoaspartyl O-methyltransferase (PIMT, EC 2.1.1.77) is a repairing enzyme converting L-isoaspartyl/D-aspartyl residues in damaged proteins to normal L-aspartyl residues. In this study it was investigated, whether PIMT is able to modulate Abeta fibrillogenesis in vitro by methylation of isoaspartyl residue using purified 5Abeta and PIMT. A Thioflavin-T (Th-T) binding assay conducted after aging Abeta in vitro (37 degrees C, pH 7.4 in PBS) revealed that PIMT inhibited the increase of fluorescence caused by amyloid fibrillogenesis. Western blot analysis revealed that high molecular Abeta aggregates (> 200 kDa) only occurred during Abeta incubation, while they were reduced in response to incubation with PIMT and AdoMet. Additionally, circular dichroism (CD) showed that the beta-sheet structure was increased in Abeta peptides in a time-dependent fashion, while PIMT suppressed the beta-sheet transition after 24 h. Finally, transmission electron microscopy (TEM) revealed that PIMT reduced the size of the Abeta aggregates and induced a different pathway, leading to the formation of amorphous structures. Taken together, these findings indicate that isoaspartyl methylation leads to partial blockade of fibrillogenesis of Abeta by inhibiting the beta transition in the Abeta peptide.
authors.affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon, Korea.
authors.forename: Gangsoo
authors.initials: G
authors.lastname: Jung
authors.forename: Jiyeon
authors.initials: J
authors.lastname: Ryu
authors.forename: Jieun
authors.initials: J
authors.lastname: Heo
authors.forename: Sang-Jin
authors.initials: SJ
authors.lastname: Lee
authors.forename: Jae Youl
authors.initials: JY
authors.lastname: Cho
authors.forename: Sungyoul
authors.initials: S
authors.lastname: Hong
mesh_terms: D016229:Amyloid beta-Peptides / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D052160:Benzothiazoles; D015153:Blotting, Western; D002942:Circular Dichroism; D004926:Escherichia coli / Q000737:chemistry; D005456:Fluorescent Dyes; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008970:Molecular Weight; D058225:Plaque, Amyloid / Q000737:chemistry*; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase / Q000302:isolation & purification / Q000494:pharmacology*; D017433:Protein Structure, Secondary; D013844:Thiazoles / Q000737:chemistry
chemical_list: D016229:Amyloid beta-Peptides; D052160:Benzothiazoles; D005456:Fluorescent Dyes; D013844:Thiazoles; C009462:thioflavin T; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase