Conformational analysis of misfolded protein aggregation by FRET and live-cell imaging techniques

International Journal of Molecular Sciences
Akira KitamuraMasataka Kinjo

Abstract

Cellular homeostasis is maintained by several types of protein machinery, including molecular chaperones and proteolysis systems. Dysregulation of the proteome disrupts homeostasis in cells, tissues, and the organism as a whole, and has been hypothesized to cause neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS) and Huntington's disease (HD). A hallmark of neurodegenerative disorders is formation of ubiquitin-positive inclusion bodies in neurons, suggesting that the aggregation process of misfolded proteins changes during disease progression. Hence, high-throughput determination of soluble oligomers during the aggregation process, as well as the conformation of sequestered proteins in inclusion bodies, is essential for elucidation of physiological regulation mechanism and drug discovery in this field. To elucidate the interaction, accumulation, and conformation of aggregation-prone proteins, in situ spectroscopic imaging techniques, such as Förster/fluorescence resonance energy transfer (FRET), fluorescence correlation spectroscopy (FCS), and bimolecular fluorescence complementation (BiFC) have been employed. Here, we summarize recent reports in which these techniques were applied to the analysis of agg...Continue Reading

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Citations

May 13, 2017·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Akira KitamuraMasataka Kinjo
Mar 24, 2018·International Journal of Molecular Sciences·Akira Kitamura, Masataka Kinjo
Apr 16, 2020·Nanoscale·Candace E BenjaminJeremiah J Gassensmith
Jun 20, 2017·Journal of Peptide Science : an Official Publication of the European Peptide Society·Kenichi KawanoKatsumi Matsuzaki
May 6, 2021·Biochemical and Biophysical Research Communications·Akira KitamuraMasataka Kinjo
Jun 9, 2018·Journal of the American Chemical Society·Yu LiuXin Zhang
Sep 12, 2020·Journal of the American Chemical Society·Charles H WolstenholmeXin Zhang
Oct 7, 2021·Chembiochem : a European Journal of Chemical Biology·Di ShenYu Liu

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Methods Mentioned

BETA
FRET
FCS
fluorescence complementation
Fluorescence
X-ray
NMR
biosensors

Software Mentioned

FCCS
BiFC

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