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A Method to Quantify Molecular Diffusion within Thin Solvated Polymer Films: A Case Study on Films of Natively Unfolded Nucleoporins
[Image: see text] 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 po...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American
Chemical Society
2020
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7526988/ https://www.ncbi.nlm.nih.gov/pubmed/32667780 http://dx.doi.org/10.1021/acsnano.0c02895 |
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author | Frost, Rickard Débarre, Delphine Jana, Saikat Bano, Fouzia Schünemann, Jürgen Görlich, Dirk Richter, Ralf P. |
author_facet | Frost, Rickard Débarre, Delphine Jana, Saikat Bano, Fouzia Schünemann, Jürgen Görlich, Dirk Richter, Ralf P. |
author_sort | Frost, Rickard |
collection | PubMed |
description | [Image: see text] 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. |
format | Online Article Text |
id | pubmed-7526988 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | American
Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-75269882020-10-01 A Method to Quantify Molecular Diffusion within Thin Solvated Polymer Films: A Case Study on Films of Natively Unfolded Nucleoporins Frost, Rickard Débarre, Delphine Jana, Saikat Bano, Fouzia Schünemann, Jürgen Görlich, Dirk Richter, Ralf P. ACS Nano [Image: see text] 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. American Chemical Society 2020-07-15 2020-08-25 /pmc/articles/PMC7526988/ /pubmed/32667780 http://dx.doi.org/10.1021/acsnano.0c02895 Text en This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. |
spellingShingle | Frost, Rickard Débarre, Delphine Jana, Saikat Bano, Fouzia Schünemann, Jürgen Görlich, Dirk Richter, Ralf P. A Method to Quantify Molecular Diffusion within Thin Solvated Polymer Films: A Case Study on Films of Natively Unfolded Nucleoporins |
title | A Method to Quantify Molecular Diffusion within Thin
Solvated Polymer Films: A Case Study on Films of Natively Unfolded
Nucleoporins |
title_full | A Method to Quantify Molecular Diffusion within Thin
Solvated Polymer Films: A Case Study on Films of Natively Unfolded
Nucleoporins |
title_fullStr | A Method to Quantify Molecular Diffusion within Thin
Solvated Polymer Films: A Case Study on Films of Natively Unfolded
Nucleoporins |
title_full_unstemmed | A Method to Quantify Molecular Diffusion within Thin
Solvated Polymer Films: A Case Study on Films of Natively Unfolded
Nucleoporins |
title_short | A Method to Quantify Molecular Diffusion within Thin
Solvated Polymer Films: A Case Study on Films of Natively Unfolded
Nucleoporins |
title_sort | method to quantify molecular diffusion within thin
solvated polymer films: a case study on films of natively unfolded
nucleoporins |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7526988/ https://www.ncbi.nlm.nih.gov/pubmed/32667780 http://dx.doi.org/10.1021/acsnano.0c02895 |
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