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| Type |
Poster Presentation |
| Area |
Life Chemistry |
| Room No. |
Exhibition Hall 2 |
| Time |
4월 19일 (금요일) 11:00~12:30 |
| Code |
LIFE.P-369 |
| Subject |
Development of a real-time FRET detection system for amyloid-β plaque formation towards therapeutic treatment of Alzheimer disease. |
| Authors |
Sang Jeon Chung*, Yeon Gyo Hwang1
College of Pharmacy, SungKyunKwan University, Korea
1Pharmacy, Sungkyunkwan University, Korea
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| Abstract |
Alzheimer disease (AD) occurs by aggregation of amyloid-β and its accumulation of amyloid-β plaque in the brain can be fatal. Amyloid-β is formed by the cleavage of the amyloid protein precursor (APP). Aggregation of amyloid-β is highly influenced by high concentrations of metal (II) ions in the brain. These metal ions coordinate especially with histidine, glutamic acid, and aspartic acid residues of amyloid-β.
There are several methods to detect amyloid-β aggregation, for example, the use of nuclear magnetic resonance (NMR) and transmission electron microscopy (TEM). However, these techniques can be laborious, expensive, and often requires dedicated instruments.
To overcome the drawbacks of the aforementioned methods, we designed a fast and reliable system to detect amyloid-β aggregation based on Förster resonance energy transfer (FRET). Real-time imaging of amyloid-β aggregation was achieved using an amyloid-β fragment equipped with an unnatural fluorescent amino acid. But using dye with short wavelength range, interference may occur, so we used dye which are long wavelength band. In our design, a C-terminal FITC residue of the amyloid-β fragment act as the FRET donor to the Rhodamine b residue. However, only upon aggregation of the construct, the FRET pair is in close proximity to allow energy transfer.
Consequently, excitation at 490 nm (i.e., FITC excitation maximum) results in significant sensitized emission at 580 nm (i.e., Rhodamine b emission maximum) upon addition of a metal ion to the artificial amyloid-β fragment. Our FRET based approach allows monitoring of the metal ion induced amyloid-β plaque formation. The developed method may be applicable in the high throughput screening of potential inhibitors of amyloid-β aggregation for the therapeutic treatment of AD. |
| E-mail |
dusry411@naver.com |
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123rd General Meeting of the KCS