کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1172991 1491349 2016 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Nuclear magnetic resonance evidence for the dimer formation of beta amyloid peptide 1–42 in 1,1,1,3,3,3-hexafluoro-2-propanol
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Nuclear magnetic resonance evidence for the dimer formation of beta amyloid peptide 1–42 in 1,1,1,3,3,3-hexafluoro-2-propanol
چکیده انگلیسی

Alzheimer's disease involves accumulation of senile plaques in which filamentous aggregates of amyloid beta (Aβ) peptides are deposited. Recent studies demonstrate that oligomerization pathways of Aβ peptides may be complicated. To understand the mechanisms of Aβ(1–42) oligomer formation in more detail, we have established a method to produce 15N-labeled Aβ(1–42) suited for nuclear magnetic resonance (NMR) studies. For physicochemical studies, the starting protein material should be solely monomeric and all Aβ aggregates must be removed. Here, we succeeded in fractionating a “precipitation-resistant” fraction of Aβ(1–42) from an “aggregation-prone” fraction by high-performance liquid chromatography (HPLC), even from bacterially overexpressed Aβ(1–42). However, both Aβ(1–42) fractions after 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) treatment formed amyloid fibrils. This indicates that the “aggregation seed” was not completely monomerized during HFIP treatment. In addition, Aβ(1–42) dissolved in HFIP was found to display a monomer–dimer equilibrium, as shown by two-dimensional 1H–15N NMR. We demonstrated that the initial concentration of Aβ during the HFIP pretreatment altered the kinetic profiles of Aβ fibril formation in a thioflavin T fluorescence assay. The findings described here should ensure reproducible results when studying the Aβ(1–42) peptide.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 498, 1 April 2016, Pages 59–67
نویسندگان
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