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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Life Science / Faculty of Advanced Life Science >
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Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
| Title: | Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy |
| Authors: | Kitamura, Akira Browse this author →KAKEN DB | | Fujimoto, Ai Browse this author | | Kinjo, Masataka Browse this author |
| Issue Date: | Apr-2021 |
| Publisher: | Journal of visualized experiments |
| Journal Title: | JoVE-journal of visualized experiments |
| Volume: | 170 |
| Start Page: | e62576 |
| Publisher DOI: | 10.3791/62576 |
| Abstract: | Protein aggregation is a hallmark of neurodegenerative disorders such as amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), and so on. To detect and analyze soluble or diffuse protein oligomers or aggregates, fluorescence correlation spectroscopy (FCS), which can detect the diffusion speed and brightness of a single particle with a single molecule sensitivity, has been used. However, the proper procedure and know-how for protein aggregation detection have not been widely shared. Here, we show a standard procedure of FCS measurement for diffusion properties of aggregation-prone proteins in cell lysate and live cells: ALS-associated 25 kDa carboxyl-terminal fragment of TAR DNA/RNA-binding protein 43 kDa (TDP25) and superoxide dismutase 1 (SOD1). The representative results show that a part of aggregates of green fluorescent protein (GFP)-tagged TDP25 was slightly included in the soluble fraction of murine neuroblastoma Neuro2a cell lysate. Moreover, GFP-tagged SOD1 carrying ALS-associated mutation shows a slower diffusion in live cells. Accordingly, we here introduce the procedure to detect the protein aggregation via its diffusion property using FCS. |
| Type: | article |
| URI: | http://hdl.handle.net/2115/82099 |
| Appears in Collections: | 生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 北村 朗
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