Beads Screening Method of Inhibitors to Amyloid--Protein Aggregation
Abstract:We have established a simple and fast screening method of potent inhibitors to amyloid--protein (A) aggregation using a new beads technology. It is known that fibril formation of A obeys the nucleation dependent polymerization model in which small nuclei are slowly formed and fast fibril formation of A proceeds subsequently. Congo red and thioflavin T bind in specific and regular fashions to amyloid fibrils possessing -sheet structures, which results in spectral changes and fluorescence emission, respectively. Therefore, these compounds have been used for determination of amyloid fibril formation. However, there is a long delay time before the fibril formation starts, because the formation of nuclei is very slow. This means that it takes a long time to evaluate inhibitors of fibril formation in the conventional methods using Congo red and thioflavin T. In contrast to these conventional methods, the present new method using beads is thought to detect the early stage of A aggregation, since the A-immobilized beads can easily coagulate by cross-linking with free A in solution. In this regard, the present beads method is superior to the conventional ones. We have also revealed that A (1–40) and A (1–42) strictly recognize the individual structure.
Document Type: Research Article
Publication date: September 1, 2005
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