4.8 Article

TUBright: A Peptide Probe for Imaging Microtubules

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ANALYTICAL CHEMISTRY
卷 -, 期 -, 页码 -

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AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.2c01285

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资金

  1. National Key R&D Program of China [2017YFA0503502]
  2. National Natural Science Foundation of China [31730050, 32070708, 31991193, 31922018]
  3. IDG/McGovern Institute for Brain Research at Tsinghua University

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This study developed a peptide probe called TUBright that can label in vitro reconstituted microtubules with high signal-to-noise ratio. TUBright does not interfere with the dynamic behaviors of microtubules and microtubule-associated proteins.
In vitro assays using reconstituted microtubules have provided molecular insights into the principles of microtubule dynamics and the roles of microtubule-associated proteins. Emerging questions that further uncover the complexity in microtubule dynamics, especially those on tubulin isotypes and post-translational modifications, raise new technical challenges on how to visualize microtubules composed of tubulin purified from limited sources, primarily due to the low efficiency of the conventional tubulin labeling protocol. Here, we develop a peptide probe, termed TUBright, that labels in vitro reconstituted microtubules. TUBright, when coupled with different fluorescent dyes, provides flexible labeling of microtubules with a high signal-to-noise ratio. TUBright does not interfere with the dynamic behaviors of microtubules and microtubule-associated proteins. Therefore, TUBright is a useful tool for imaging microtubules, making it feasible to use tubulin from limited sources for answering many open questions on microtubule dynamics.

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