Identifying and quantifying two ligand-binding sites while imaging native human membrane receptors by AFM.

ABSTRACT: 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

SUBMITTER: Pfreundschuh M 

PROVIDER: S-EPMC4660198 | biostudies-literature | 2015 Nov

REPOSITORIES: biostudies-literature