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  A method to quantify molecular diffusion within thin solvated polymer films: A case study on films of natively unfolded nucleoporins

Frost, R., Débarre, D., Jana, S., Bano, F., Schünemann, J., Görlich, D., et al. (2020). A method to quantify molecular diffusion within thin solvated polymer films: A case study on films of natively unfolded nucleoporins. ACS Nano, 14(8), 9938-9952. doi:10.1021/acsnano.0c02895.

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Frost, R., Author
Débarre, D., Author
Jana, S., Author
Bano, F., Author
Schünemann, J.1, Author              
Görlich, D.1, Author              
Richter, R. P., Author
Affiliations:
1Department of Cellular Logistics, MPI for biophysical chemistry, Max Planck Society, ou_578574              

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Free keywords: diffusion; absorption; confinement; polymer film; reflection interference contrast microscopy; fluorescence recovery after photobleaching; permeability barrier
 Abstract: We present a method to probe molecular and nanoparticle diffusion within thin, solvated polymer coatings. The device exploits the confinement with well-defined geometry that forms at the interface between a planar and a hemispherical surface (of which at least one is coated with polymers) in close contact and uses this confinement to analyze diffusion processes without interference of exchange with and diffusion in the bulk solution. With this method, which we call plane–sphere confinement microscopy (PSCM), information regarding the partitioning of molecules between the polymer coating and the bulk liquid is also obtained. Thanks to the shape of the confined geometry, diffusion and partitioning can be mapped as a function of compression and concentration of the coating in a single experiment. The method is versatile and can be integrated with conventional optical microscopes; thus it should find widespread use in the many application areas exploiting functional polymer coatings. We demonstrate the use of PSCM using brushes of natively unfolded nucleoporin domains rich in phenylalanine–glycine repeats (FG domains). A meshwork of FG domains is known to be responsible for the selective transport of nuclear transport receptors (NTRs) and their macromolecular cargos across the nuclear envelope that separates the cytosol and the nucleus of living cells. We find that the selectivity of NTR uptake by FG domain films depends sensitively on FG domain concentration and that the interaction of NTRs with FG domains obstructs NTR movement only moderately. These observations contribute important information to better understand the mechanisms of selective NTR transport.

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Language(s): eng - English
 Dates: 2020-07-152020-08-25
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1021/acsnano.0c02895
 Degree: -

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Title: ACS Nano
Source Genre: Journal
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Pages: - Volume / Issue: 14 (8) Sequence Number: - Start / End Page: 9938 - 9952 Identifier: -