Chemically inducible diffusion trap at cilia reveals molecular sieve-like barrier.

Chemically inducible diffusion trap at cilia reveals molecular sieve-like barrier.
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DOI:
10.1038/nchembio.1252
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发表时间:
2013-07
影响因子:
14.8
通讯作者:
Inoue, Takanari
Inoue, Takanari
中科院分区:
生物学1区
文献类型:
--
作者:
Lin, Yu-Chun;Niewiadomski, Pawel;Lin, Benjamin;Nakamura, Hideki;Phua, Siew Cheng;Jiao, John;Levchenko, Andre;Inoue, Takafumi;Rohatgi, Rajat;Inoue, Takanari

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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.
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