Untersuchungen zur molekularen Dynamik von festen Proteinen

Aiming to further clarify the role of internal protein mobility in the binding process, we are intending to perform dynamic Solid-State NMR experiments on a per-residue basis. Since the dynamic range of molecular motions spans many orders of magnitude from 10-10 to 100 s, a number of different dynamic Solid-State NMR methods need to be used. Of particular importance is the characterisation of the amplitude of the dynamic processes in the different time scales, since this parameter is believed to correlate with the biologic function. This can be done in Solid-State NMR much more accurately and over a much wider dynamic range as compared to liquid-state NMR approaches. The present project consist in the application of well-established as well as recently developed dynamic NMR experiments, which need to be complemented by recently developed approaches to ensure the site-specific resolution in the Solid-state NMR spectra. For that, the preparation of isotopically labelled samples as well the growing of microcrystals is a must. The project thus is a combination of the two large parts (i) sample preparation and (ii) establishment and application of the NMR experiments that will be conducted as much as possible in parallel.

We intent to apply this approach to the protein barstar in the free and barnase-bound states. The aims of the project are to understand the role of the conformational dynamics in the binding process and a general development of methodological approaches to investigation of biopolymer dynamics by means of various NMR techniques and thus paving the path for further application on biologically relevant samples.