Project Details
Projekt
Protein labeling in living cells - A combination of enzyme mediated peptide labeling and inverse electron demand Diels-Alder cycloaddition
Applicant
Dr. Richard Wombacher
Subject Area
Biological and Biomimetic Chemistry
Term
from 2012 to 2021
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 223427877
We recently developed a protein labeling strategy that makes significant impact with regard to selective protein labeling in live cells by providing a minimally disruptive technology (minimal tag size). This labeling strategy will now serve as key technology for a novel protein specific chemoproteomic tool which we call ProSpecT (protein specific trifunctional capture probe) for the identification of receptor-ligand interactions. Most valuable asset of the tool is its potential application in live cells, tissues or organisms which makes it highly attractive for the field of interactomics and proteomics. First, we will design and synthesize different variants of ProSpecT-probes which then will be evaluated in an in vitro assay based on a well-known receptor-ligand pair, namely erythropoietin and the soluble form of the extracellular domain of the human erythropoietin receptor. Then, in a proof of concept experiment we will show that we can capture and identify the known receptors of the human vascular endothelial growth factor (VEGF) from living cells. The ambitious final goal of our proposal is to identify unknown cell surface receptor-ligand interactions and to demonstrate that ProSpecT is a highly versatile reagent and method to serve as a novel key method for functional genomics screens. The new technology will be broadly useful to the biomedical research community and can make significant impact on our understanding of the mechanisms of biological pathways and human diseases.
DFG Programme
Priority Programmes
