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

Autor: Jatzlau, Jerome, Burdzinski, Wiktor, Trumpp, Michael, Obendorf, Leon, Roßmann, Kilian, Ravn, Katharina, Hyvönen, Marko, Bottanelli, Francesca, Broichhagen, Johannes, Knaus, Petra
Zdroj: Communications Biology; 1/12/2023, Vol. 6 Issue 1, p1-22, 22p
Abstrakt: 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.A TGFβ/BMP receptor library with Halo- and SNAP-tags is generated and with the application of labeled growth factor ligands, a ligand surface binding assay is developed, allowing assessment of preferential ligand-receptor binding in cells. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje