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A versatile Halo- and SNAP-tagged BMP/TGFβ receptor library for quantification of cell surface ligand binding

TGFβs, BMPs and Activins regulate numerous developmental and homeostatic processes and signal through hetero-tetrameric receptor complexes composed of two types of serine/threonine kinase receptors. Each of the 33 different ligands possesses unique affinities towards specific receptor types. However...

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Autores principales: Jatzlau, Jerome, Burdzinski, Wiktor, Trumpp, Michael, Obendorf, Leon, Roßmann, Kilian, Ravn, Katharina, Hyvönen, Marko, Bottanelli, Francesca, Broichhagen, Johannes, Knaus, Petra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9837045/
https://www.ncbi.nlm.nih.gov/pubmed/36635368
http://dx.doi.org/10.1038/s42003-022-04388-4
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author Jatzlau, Jerome
Burdzinski, Wiktor
Trumpp, Michael
Obendorf, Leon
Roßmann, Kilian
Ravn, Katharina
Hyvönen, Marko
Bottanelli, Francesca
Broichhagen, Johannes
Knaus, Petra
author_facet Jatzlau, Jerome
Burdzinski, Wiktor
Trumpp, Michael
Obendorf, Leon
Roßmann, Kilian
Ravn, Katharina
Hyvönen, Marko
Bottanelli, Francesca
Broichhagen, Johannes
Knaus, Petra
author_sort Jatzlau, Jerome
collection PubMed
description TGFβs, BMPs and Activins regulate numerous developmental and homeostatic processes and signal through hetero-tetrameric receptor complexes composed of two types of serine/threonine kinase receptors. Each of the 33 different ligands possesses unique affinities towards specific receptor types. However, the lack of specific tools hampered simultaneous testing of ligand binding towards all BMP/TGFβ receptors. Here we present a N-terminally Halo- and SNAP-tagged TGFβ/BMP receptor library to visualize receptor complexes in dual color. In combination with fluorescently labeled ligands, we established a Ligand Surface Binding Assay (LSBA) for optical quantification of receptor-dependent ligand binding in a cellular context. We highlight that LSBA is generally applicable to test (i) binding of different ligands such as Activin A, TGFβ1 and BMP9, (ii) for mutant screens and (iii) evolutionary comparisons. This experimental set-up opens opportunities for visualizing ligand-receptor binding dynamics, essential to determine signaling specificity and is easily adaptable for other receptor signaling pathways.
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spelling pubmed-98370452023-01-14 A versatile Halo- and SNAP-tagged BMP/TGFβ receptor library for quantification of cell surface ligand binding Jatzlau, Jerome Burdzinski, Wiktor Trumpp, Michael Obendorf, Leon Roßmann, Kilian Ravn, Katharina Hyvönen, Marko Bottanelli, Francesca Broichhagen, Johannes Knaus, Petra Commun Biol Article TGFβs, BMPs and Activins regulate numerous developmental and homeostatic processes and signal through hetero-tetrameric receptor complexes composed of two types of serine/threonine kinase receptors. Each of the 33 different ligands possesses unique affinities towards specific receptor types. However, the lack of specific tools hampered simultaneous testing of ligand binding towards all BMP/TGFβ receptors. Here we present a N-terminally Halo- and SNAP-tagged TGFβ/BMP receptor library to visualize receptor complexes in dual color. In combination with fluorescently labeled ligands, we established a Ligand Surface Binding Assay (LSBA) for optical quantification of receptor-dependent ligand binding in a cellular context. We highlight that LSBA is generally applicable to test (i) binding of different ligands such as Activin A, TGFβ1 and BMP9, (ii) for mutant screens and (iii) evolutionary comparisons. This experimental set-up opens opportunities for visualizing ligand-receptor binding dynamics, essential to determine signaling specificity and is easily adaptable for other receptor signaling pathways. Nature Publishing Group UK 2023-01-12 /pmc/articles/PMC9837045/ /pubmed/36635368 http://dx.doi.org/10.1038/s42003-022-04388-4 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Jatzlau, Jerome
Burdzinski, Wiktor
Trumpp, Michael
Obendorf, Leon
Roßmann, Kilian
Ravn, Katharina
Hyvönen, Marko
Bottanelli, Francesca
Broichhagen, Johannes
Knaus, Petra
A versatile Halo- and SNAP-tagged BMP/TGFβ receptor library for quantification of cell surface ligand binding
title A versatile Halo- and SNAP-tagged BMP/TGFβ receptor library for quantification of cell surface ligand binding
title_full A versatile Halo- and SNAP-tagged BMP/TGFβ receptor library for quantification of cell surface ligand binding
title_fullStr A versatile Halo- and SNAP-tagged BMP/TGFβ receptor library for quantification of cell surface ligand binding
title_full_unstemmed A versatile Halo- and SNAP-tagged BMP/TGFβ receptor library for quantification of cell surface ligand binding
title_short A versatile Halo- and SNAP-tagged BMP/TGFβ receptor library for quantification of cell surface ligand binding
title_sort versatile halo- and snap-tagged bmp/tgfβ receptor library for quantification of cell surface ligand binding
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9837045/
https://www.ncbi.nlm.nih.gov/pubmed/36635368
http://dx.doi.org/10.1038/s42003-022-04388-4
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