TY  - JOUR
A1  - Pfreundschuh, Moritz
A1  - Alsteens, David
A1  - Wieneke, Ralph
A1  - Zhang, Cheng
A1  - Coughlin, Shaun R.
A1  - Tampé, Robert
A1  - Kobilka, Brian K.
A1  - Müller, Daniel J.
T1  - Identifying and quantifying two ligand-binding sites while imaging native human membrane receptors by AFM
T2  - Nature Communications
N2  - A current challenge in life sciences is to image cell membrane receptors while characterizing their specific interactions with various ligands. Addressing this issue has been hampered by the lack of suitable nanoscopic methods. Here we address this challenge and introduce multifunctional high-resolution atomic force microscopy (AFM) to image human protease-activated receptors (PAR1) in the functionally important lipid membrane and to simultaneously localize and quantify their binding to two different ligands. Therefore, we introduce the surface chemistry to bifunctionalize AFM tips with the native receptor-activating peptide and a tris-N-nitrilotriacetic acid (tris-NTA) group binding to a His10-tag engineered to PAR1. We further introduce ways to discern between the binding of both ligands to different receptor sites while imaging native PAR1s. Surface chemistry and nanoscopic method are applicable to a range of biological systems in vitro and in vivo and to concurrently detect and localize multiple ligand-binding sites at single receptor resolution.
KW  - Atomic force microscopy
KW  - G protein-coupled receptors
KW  - Membrane biophysics
Y1  - 2015
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/50606
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-506062
SN  - 2041-1723
N1  - This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
VL  - 6
IS  - Art. 8857
SP  - 1
EP  - 7
PB  - Nature Publishing Group UK
CY  - [London]
ER  -