Chemically inducible diffusion trap at cilia reveals molecular sieve–like barrier
| Autor: | Andre Levchenko, Benjamin Lin, John Jiao, Rajat Rohatgi, Pawel Niewiadomski, Takanari Inoue, Hideki Nakamura, Siew Cheng Phua, Yu Chun Lin, Takafumi Inoue |
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| Rok vydání: | 2013 |
| Předmět: |
Green Fluorescent Proteins
Biology Microtubules Article Diffusion Cell membrane Mice Cytosol Bacterial Proteins Microtubule medicine Animals Humans Cilia Diffusion (business) Molecular Biology Fluorescent Dyes Luminescent Proteins Cilium Cell Membrane Cell Biology Cell biology HEK293 Cells Spectrometry Fluorescence medicine.anatomical_structure Models Chemical Chromatography Gel NIH 3T3 Cells Signal transduction Dimerization Function (biology) |
| Zdroj: | Nature Chemical Biology. 9:437-443 |
| ISSN: | 1552-4469 1552-4450 |
| DOI: | 10.1038/nchembio.1252 |
| Popis: | Primary cilia function as specialized compartments for signal transduction. The stereotyped structure and signaling function of cilia inextricably depend on the selective segregation of molecules in cilia. However, the fundamental principles governing the access of soluble proteins to primary cilia remain unresolved. We developed a methodology termed Chemically-Inducible Diffusion Trap at Cilia (C-IDTc) to visualize the diffusion process of a series of fluorescent proteins ranging in size from 3.2 to 7.9 nm into primary cilia. We found that the interior of the cilium was accessible to proteins as large as 7.9 nm. The kinetics of ciliary accumulation of this panel of proteins was exponentially limited by their Stokes radii. Quantitative modeling suggests that the diffusion barrier operates as a molecular sieve at the base of cilia. Our study presents a set of powerful, generally applicable tools for the quantitative monitoring of ciliary protein diffusion under both physiological and pathological conditions. |
| Databáze: | OpenAIRE |
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