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Cited by 20TGF-β signaling in diseasesDevelopmental Biology and Gene RegulationBiotin and Related Studies

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

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Jatzlau, Jerome, Burdzinski, Wiktor, Trumpp, Michael, Obendorf, Leon, Roßmann, Kilian, Ravn, Katharina, Hyvönen, Marko, Bottanelli, Francesca, Broichhagen, Johannes, Knaus, Petra·2023·Communications Biology
DOI 10.1038/s42003-022-04388-4PMC PMC9837045

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Abstract

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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Metadata

Authors
Jatzlau, J., Burdzinski, W., Trumpp, M., Obendorf, L., Roßmann, K., Ravn, K., Hyvönen, M., Bottanelli, F., Broichhagen, J., & Knaus, P.
Affiliations
  • Leibniz-Forschungsinstitut für Molekulare Pharmakologie (DE)
  • University of Cambridge (GB)
  • Berlin-Brandenburger Centrum für Regenerative Therapien (DE)
Corresponding author
Knaus, Petra
ORCID
Venue
Communications Biology
Publisher
Springer Science and Business Media LLC
Published
2023-01-12
Volume
6
Issue
1
Pages
34-34
ISSN
2399-3642 (ISSN-L)
DOI
10.1038/s42003-022-04388-4
PMC
PMC9837045
OpenAlex
W4315779629
Cited by
20
References
2
Citations by year
2026: 7 · 2025: 5 · 2024: 2 · 2023: 4
FWCI
1.60
Type
journal article
Language
en
Open access
Licence
Locations
  • Communications Biology (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Refubium (Universitätsbibliothek der Freien Universität Berlin) (submittedVersion, cc-by)
  • Apollo (University of Cambridge) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Universitätsbibliothek der FU Berlin Hochschulschriftenstelle u. Dokumentenserver (cc-by)
  • Apollo
  • Apollo (cc-by)
Funders
Morbus Osler Society Germany Einstein Center for Regenerative Therapies, Berlin
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › TGF-β signaling in diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
Keywords
ligand-receptor binding, HaloTag, SNAP-tag, TGF-beta signaling, activin A, BMP9, TGF-β1, receptor signaling specificity, mutant screening, receptor evolution
MeSH terms
Ligands, Transforming Growth Factor beta, Receptors, Transforming Growth Factor beta, Bone Morphogenetic Proteins, Bone Morphogenetic Protein Receptors

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