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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...

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Autores principales: Sedlak, Steffen M., Bauer, Magnus S., Kluger, Carleen, Schendel, Leonard C., Milles, Lukas F., Pippig, Diana A., Gaub, Hermann E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
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.
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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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