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

Takanari Inoue, Yu Chun Lin, Pawel Niewiadomski, Benjamin Lin, Hideki Nakamura, Siew Cheng Phua, John Jiao, Andre Levchenko, Takafumi Inoue, Rajat Rohatgi

Research output: Contribution to journalArticlepeer-review

72 Citations (Scopus)

Abstract

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' to visualize the diffusion process of a series of fluorescent proteins ranging in size from 3.2 nm 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

Original languageEnglish
Pages (from-to)437-443
Number of pages7
JournalNature Chemical Biology
Volume9
Issue number7
DOIs
Publication statusPublished - 2013 Jul 1

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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