Purification tags markedly affect self‐aggregation of <scp>CPEB3</scp>
Since protein aggregation-including liquid-liquid phase separation (LLPS) and amyloid fibril formation-plays a critical role in both diseases and biological functions, understanding the mechanisms underlying protein aggregation is essential. Recombinant proteins are commonly used in vitro to investigate protein aggregation processes. However, if the purification tags remain uncleaved, they may affect the results and hinder accurate interpretation. Our findings demonstrate that the His<sub>6</sub>-GFP and His<sub>12</sub> tags significantly affect liquid droplet and amyloid fibril formation in the intrinsically disordered region (IDR) of mouse cytoplasmic polyadenylation element-binding protein 3 (CPEB3) and its fragments. This study shows that the purification tags significantly affect aggregation assays, making it essential to account for their influence to accurately interpret protein aggregation.
Authors
- Naotaka Sekiyama (ORCID: https://orcid.org/0000-0003-4537-6565)
- Masato Ueno (ORCID: https://orcid.org/0009-0007-2410-1032)
- Masatomo So (ORCID: https://orcid.org/0000-0002-4815-4995)
- Kenji Sugase (ORCID: https://orcid.org/0000-0001-8623-7743)
- Yujin Lee (ORCID: https://orcid.org/0000-0003-0028-9476)
- Ayako Furukawa (ORCID: https://orcid.org/0000-0001-9310-9918)
- Hideya Saito
Institutions
- Kyoto University (JP)
Publication Details
- Journal
- FEBS Letters
- Published
- 2025-06-17
- DOI
- https://doi.org/10.1002/1873-3468.70090
- Primary Topic
- RNA Research and Splicing
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Japan Society for the Promotion of Science
- Institute for Protein Research, Osaka University