Probing Enzyme-Protein Interaction with Force Spectroscopy

Matthias Hofmann

Max-Planck-Institute for Biochemistry, Am Klopferspitz 18a, D-82152 Planegg-Martinsried, Germany

e-mail: mhofmann@mpg.biochem.de
 

The goal of our project is to examine the structural dynamics and mechanical details of the process of degradation of unfolded proteins by the 20 S Proteasome. In my talk I will introduce the AFM part of the project. The AFM-technique allows us to litterally watch single biomolecules at their work. Firstly high resolution and time resolved imaging of the proteasome is perfomed. Secondly single molecule force measurements will shed light on the mechanism of binding of proteins and polypeptides to 20 S, translocation into the nanocompartment and degradation itself. Prerequiries for this endeavour are the oriented deposition of the proteasome on a biocompatible surface and the functionalization of the cantilever tip with polypeptides and proteins. Several techniques will be introduced and first results wil be shown.