Journal
NATURE CHEMICAL BIOLOGY
Volume 9, Issue 7, Pages 437-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NCHEMBIO.1252
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Funding
- US National Institutes of Health (the Baltimore Polycystic Kidney Disease Research and Clinical Core Center) [GM092930, P30 DK090868, R00CA129174, R21NS074091, 23650197]
- US National Institutes of Health (Pew Foundation)
- Grants-in-Aid for Scientific Research [22650066, 23650197, 23300121] Funding Source: KAKEN
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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' 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.
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