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Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry
The widely used interaction of the homotetramer streptavidin with the small molecule biotin has been intensively studied by force spectroscopy and has become a model system for receptor ligand interaction. However, streptavidin’s tetravalency results in diverse force propagation pathways through the...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5716544/ https://www.ncbi.nlm.nih.gov/pubmed/29206886 http://dx.doi.org/10.1371/journal.pone.0188722 |
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author | Sedlak, Steffen M. Bauer, Magnus S. Kluger, Carleen Schendel, Leonard C. Milles, Lukas F. Pippig, Diana A. Gaub, Hermann E. |
author_facet | Sedlak, Steffen M. Bauer, Magnus S. Kluger, Carleen Schendel, Leonard C. Milles, Lukas F. Pippig, Diana A. Gaub, Hermann E. |
author_sort | Sedlak, Steffen M. |
collection | PubMed |
description | The widely used interaction of the homotetramer streptavidin with the small molecule biotin has been intensively studied by force spectroscopy and has become a model system for receptor ligand interaction. However, streptavidin’s tetravalency results in diverse force propagation pathways through the different binding interfaces. This multiplicity gives rise to polydisperse force spectroscopy data. Here, we present an engineered monovalent streptavidin tetramer with a single cysteine in its functional subunit that allows for site-specific immobilization of the molecule, orthogonal to biotin binding. Functionality of streptavidin and its binding properties for biotin remain unaffected. We thus created a stable and reliable molecular anchor with a unique high-affinity binding site for biotinylated molecules or nanoparticles, which we expect to be useful for many single-molecule applications. To characterize the mechanical properties of the bond between biotin and our monovalent streptavidin, we performed force spectroscopy experiments using an atomic force microscope. We were able to conduct measurements at the single-molecule level with 1:1-stoichiometry and a well-defined geometry, in which force exclusively propagates through a single subunit of the streptavidin tetramer. For different force loading rates, we obtained narrow force distributions of the bond rupture forces ranging from 200 pN at 1,500 pN/s to 230 pN at 110,000 pN/s. The data are in very good agreement with the standard Bell-Evans model with a single potential barrier at Δx(0) = 0.38 nm and a zero-force off-rate k(off,0) in the 10(−6) s(-1) range. |
format | Online Article Text |
id | pubmed-5716544 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-57165442017-12-15 Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry Sedlak, Steffen M. Bauer, Magnus S. Kluger, Carleen Schendel, Leonard C. Milles, Lukas F. Pippig, Diana A. Gaub, Hermann E. PLoS One Research Article The widely used interaction of the homotetramer streptavidin with the small molecule biotin has been intensively studied by force spectroscopy and has become a model system for receptor ligand interaction. However, streptavidin’s tetravalency results in diverse force propagation pathways through the different binding interfaces. This multiplicity gives rise to polydisperse force spectroscopy data. Here, we present an engineered monovalent streptavidin tetramer with a single cysteine in its functional subunit that allows for site-specific immobilization of the molecule, orthogonal to biotin binding. Functionality of streptavidin and its binding properties for biotin remain unaffected. We thus created a stable and reliable molecular anchor with a unique high-affinity binding site for biotinylated molecules or nanoparticles, which we expect to be useful for many single-molecule applications. To characterize the mechanical properties of the bond between biotin and our monovalent streptavidin, we performed force spectroscopy experiments using an atomic force microscope. We were able to conduct measurements at the single-molecule level with 1:1-stoichiometry and a well-defined geometry, in which force exclusively propagates through a single subunit of the streptavidin tetramer. For different force loading rates, we obtained narrow force distributions of the bond rupture forces ranging from 200 pN at 1,500 pN/s to 230 pN at 110,000 pN/s. The data are in very good agreement with the standard Bell-Evans model with a single potential barrier at Δx(0) = 0.38 nm and a zero-force off-rate k(off,0) in the 10(−6) s(-1) range. Public Library of Science 2017-12-05 /pmc/articles/PMC5716544/ /pubmed/29206886 http://dx.doi.org/10.1371/journal.pone.0188722 Text en © 2017 Sedlak et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Sedlak, Steffen M. Bauer, Magnus S. Kluger, Carleen Schendel, Leonard C. Milles, Lukas F. Pippig, Diana A. Gaub, Hermann E. Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry |
title | Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry |
title_full | Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry |
title_fullStr | Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry |
title_full_unstemmed | Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry |
title_short | Monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry |
title_sort | monodisperse measurement of the biotin-streptavidin interaction strength in a well-defined pulling geometry |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5716544/ https://www.ncbi.nlm.nih.gov/pubmed/29206886 http://dx.doi.org/10.1371/journal.pone.0188722 |
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