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Artificial Intelligence-Enabled AlphaFold II Pipeline Guides Functional Fluorescence Labeling of Tubulin Across Species
Open accessPreprint — published as 10.1371/journal.pbio.3002615 · Paper page
Abstract
Dynamic properties are essential for microtubule (MT) physiology. Current techniques for in vivo imaging of MTs present intrinsic limitations in elucidating the isotype-specific nuances of tubulins, which contribute to their versatile functions. Harnessing the power of AlphaFold II pipeline, we engineered a strategy for the minimally invasive fluorescence labeling of endogenous tubulin isotypes or those harboring missense mutations. We demonstrated that a specifically designed 16-amino acid linker, coupled with sfGFP11 from the split-sfGFP system and integration into the H1-S2 loop of tubulin, facilitated tubulin labeling without compromising MT dynamics, embryonic development, or ciliogenesis in C. elegans . Extending this technique to human cells and murine oocytes, we visualized MTs with the minimal background fluorescence and a pathogenic tubulin isoform with fidelity. The utility of our approach across biological contexts and species set an additional paradigm for studying tubulin dynamics and functional specificity, with implications for understanding tubulin-related diseases known as tubulinopathies.
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Highly accurate protein structure prediction with AlphaFold
Jumper, J., Evans, R., Pritzel, A., Green, T., Figurnov, M., Ronneberger, O., Tunyasuvunakool, K., Bates, R., Žídek, A., Potapenko, A., et al. (2021) · Nature · doi:10.1038/s41586-021-03819-2 · Cited by 43,764
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Metadata
- Authors
- Xu, K., Li, Z., Mao, L., Guo, Z., Chen, Z., Chai, Y., Xie, C., Yang, X., Na, J., Li, W., & Ou, G.
- Affiliations
- McGovern Institute for Brain Research (US)
- Tsinghua University (CN)
- Corresponding author
- Xu, Kaiming
- ORCID
- Venue
- PLoS Biology
- Publisher
- openRxiv
- Published
- 2024-04-11
- Volume
- 22
- Issue
- 8
- Pages
- e3002615-e3002615
- ISSN
- 1544-9173 (ISSN-L)
- PMC
- PMC11361732
- OpenAlex
- W4401690942
- Cited by
- 21
- References
- 1
- Citations by year
- 2026: 5 · 2025: 15 · 2024: 1
- FWCI
- 4.70
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- PLoS Biology (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- U.S. Department of Veterans Affairs (I01 HX002025)
- National Natural Science Foundation of China (31671444, 31525015, 31991190, 31730052, 31525015, 31861143042, 31561130153, 31671444, and 31871352, 31730052, 31861143042, 31561130153, 323B200173, 2017YFA0102900, and 31871352, 31871352, 31991190)
- Tsinghua University
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
- Keywords
- split GFP, microtubules, tubulin isoforms, missense mutations, Caenorhabditis elegans, ciliogenesis, tubulinopathy, microtubule dynamics
- MeSH terms
- Animals, Humans, Microtubules, Oocytes, Tubulin, Protein Engineering, Caenorhabditis elegans, Green Fluorescent Proteins, Mice
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