citation_mjid: advances;5/8/eaaw4988 og-title: Dynamic tuning of FRET in a green fluorescent protein biosensor og:image: https://advances.sciencemag.org/content/5/8/F1.medium.gif article:published_time: 2019-08-01 twitter:card: summary_large_image citation_reference: citation_journal_title=Chemical Reviews;citation_journal_abbrev=Chemical Reviews;citation_author=R. H. Newman;citation_author=M. D. Fosbrink;citation_author=J. Zhang;citation_title=Genetically encodable fluorescent biosensors for tracking signaling dynamics in living cells.;citation_pages=3614-3666;citation_volume=111;citation_year=2011;citation_issue=5;citation_pmid=21456512;citation_doi=10.1021/cr100002u citation_journal_title: Science Advances type: article citation_author_email: sbecker2@gwdg.de citation_issn: 2375-2548 citation_full_html_url: https://advances.sciencemag.org/content/5/8/eaaw4988.full dc:title: Dynamic tuning of FRET in a green fluorescent protein biosensor | Science Advances citation_public_url: https://advances.sciencemag.org/content/5/8/eaaw4988 Content-Encoding: UTF-8 citation_pdf_url: https://advances.sciencemag.org/content/advances/5/8/eaaw4988.full.pdf citation_section: Research Article citation_fulltext_world_readable: DC.Identifier: 10.1126/sciadv.aaw4988 DC.Rights: Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).. This is an open-access article distributed under the terms of the Creative Commons Attribution license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. citation_author: Pablo Trigo-Mourino citation_abstract_html_url: https://advances.sciencemag.org/content/5/8/eaaw4988.abstract citation_issue: 8 og-image: https://advances.sciencemag.org/sites/default/files/highwire/advances/5/8.cover-source.gif HW.identifier: /advances/5/8/eaaw4988.atom citation_doi: 10.1126/sciadv.aaw4988 citation_volume: 5 Content-Language: en Generator: Drupal 7 (http://drupal.org) citation_author_orcid: http://orcid.org/0000-0003-0822-882X DC.AccessRights: open-access citation_publication_date: 2019/08/01 citation_title: Dynamic tuning of FRET in a green fluorescent protein biosensor citation_author_institution: Department for NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany. citation_publisher: American Association for the Advancement of Science HandheldFriendly: true citation_id: 5/8/eaaw4988 og-type: article cleartype: on title: Dynamic tuning of FRET in a green fluorescent protein biosensor | Science Advances twitter:image: https://advances.sciencemag.org/sites/default/files/highwire/advances/5/8.cover-source.gif og-url: https://advances.sciencemag.org/content/5/8/eaaw4988 DC.Description: Förster resonance energy transfer (FRET) between mutants of green fluorescent protein is widely used to monitor protein-protein interactions and as a readout mode in fluorescent biosensors. Despite the fundamental importance of distance and molecular angles of fluorophores to each other, structural details on fluorescent protein FRET have been missing. Here, we report the high-resolution x-ray structure of the fluorescent proteins mCerulean3 and cpVenus within the biosensor Twitch-2B, as they undergo FRET and characterize the dynamics of this biosensor with B02-dependent paramagnetic nuclear magnetic resonance at 900 MHz and 1.1 GHz. These structural data provide the unprecedented opportunity to calculate FRET from the x-ray structure and to compare it to experimental data in solution. We find that interdomain dynamics limits the FRET effect and show that a rigidification of the sensor further enhances FRET. og-description: Förster resonance energy transfer (FRET) between mutants of green fluorescent protein is widely used to monitor protein-protein interactions and as a readout mode in fluorescent biosensors. Despite the fundamental importance of distance and molecular angles of fluorophores to each other, structural details on fluorescent protein FRET have been missing. Here, we report the high-resolution x-ray structure of the fluorescent proteins mCerulean3 and cpVenus within the biosensor Twitch-2B, as they undergo FRET and characterize the dynamics of this biosensor with B02-dependent paramagnetic nuclear magnetic resonance at 900 MHz and 1.1 GHz. These structural data provide the unprecedented opportunity to calculate FRET from the x-ray structure and to compare it to experimental data in solution. We find that interdomain dynamics limits the FRET effect and show that a rigidification of the sensor further enhances FRET. Content-Type-Hint: text/html; charset=utf-8 DC.Format: text/html DC.Publisher: American Association for the Advancement of Science DC.Contributor: Pablo Trigo-Mourino Content-Type: application/xhtml+xml; charset=UTF-8 X-Parsed-By: org.apache.tika.parser.DefaultParser article:section: Research Article twitter:title: Dynamic tuning of FRET in a green fluorescent protein biosensor citation_article_type: Research Article citation_abstract:

Förster resonance energy transfer (FRET) between mutants of green fluorescent protein is widely used to monitor protein-protein interactions and as a readout mode in fluorescent biosensors. Despite the fundamental importance of distance and molecular angles of fluorophores to each other, structural details on fluorescent protein FRET have been missing. Here, we report the high-resolution x-ray structure of the fluorescent proteins mCerulean3 and cpVenus within the biosensor Twitch-2B, as they undergo FRET and characterize the dynamics of this biosensor with B02-dependent paramagnetic nuclear magnetic resonance at 900 MHz and 1.1 GHz. These structural data provide the unprecedented opportunity to calculate FRET from the x-ray structure and to compare it to experimental data in solution. We find that interdomain dynamics limits the FRET effect and show that a rigidification of the sensor further enhances FRET.

DC.Title: Dynamic tuning of FRET in a green fluorescent protein biosensor issue_cover_image: https://advances.sciencemag.org/sites/default/files/highwire/advances/5/8.cover-source.gif citation_firstpage: eaaw4988 X-UA-Compatible: IE=edge,chrome=1 MobileOptimized: width citation_funding_source: citation_funder=Alexander von Humboldt-Stiftung;citation_funder_id=http://dx.doi.org/10.13039/100005156; HW.pisa: advances;5/8/eaaw4988 viewport: width=device-width, initial-scale=1 DC.Language: en twitter:description: Förster resonance energy transfer (FRET) between mutants of green fluorescent protein is widely used to monitor protein-protein interactions and as a readout mode in fluorescent biosensors. Despite the fundamental importance of distance and molecular angles of fluorophores to each other, structural details on fluorescent protein FRET have been missing. Here, we report the high-resolution x-ray structure of the fluorescent proteins mCerulean3 and cpVenus within the biosensor Twitch-2B, as they undergo FRET and characterize the dynamics of this biosensor with B02-dependent paramagnetic nuclear magnetic resonance at 900 MHz and 1.1 GHz. These structural data provide the unprecedented opportunity to calculate FRET from the x-ray structure and to compare it to experimental data in solution. We find that interdomain dynamics limits the FRET effect and show that a rigidification of the sensor further enhances FRET. DC.Date: 2019-08-01 citation_access: all og-site-name: Science Advances category: research-article