title: Fluorescent and luminescent fusion proteins for analyses of amyloid beta peptide aggregation.
abstract: The amyloid beta (Aβ) peptide is regarded as a causative agent of Alzheimer's disease. In this study, fluorescent and luminescent fusion proteins were constructed to analyze Aβ aggregation. A system was developed to monitor changes in luminescence that provides information about Aβ aggregation. In the presence of monomeric Aβ, the fusion protein exhibits higher luminescence intensity, and the luminescence intensity is diminished after aggregation of the fusion protein and Aβ. In contrast, the fluorescence is sustained in the presence of Aβ. In the absence of Aβ, the fusion protein self-aggregates, and its luminescence and fluorescence are quenched, thus decreasing the background fluorescence and enhancing the detection of Aβ inside and outside the cells. The ratio of the luminescence intensity to the fluorescence intensity would allow the aggregation degrees of Aβ to be distinguished. This study would be a promising method for analyzing the aggregation state of a particular amyloid protein/peptide (monomer, oligomer, or fibril), as well as the distribution of the amyloid protein/peptide within and at the cell surface, by using a single fusion protein. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.
authors.affiliation: Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, Kobe, Japan.
authors.forename: Kenji
authors.identifier: http://orcid.org/0000-0003-4558-5762
authors.initials: K
authors.lastname: Usui
authors.affiliation: School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Masayasu
authors.initials: M
authors.lastname: Mie
authors.affiliation: School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Andou
authors.affiliation: School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Mihara
authors.affiliation: School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Eiry
authors.initials: E
authors.lastname: Kobatake
mesh_terms: D016229:Amyloid beta-Peptides / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D016229:Amyloid beta-Peptides; D008164:Luminescent Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins
keywords: Alzheimer's disease; amyloid beta peptide; fluorescent protein; fusion protein; luminescent protein