Biochemistry

Main focus: Cell-free synthesis of proteins
One of the biotechnological applications in the field of proteomics that is well established in our work group is cell-free synthesis of proteins. This method (A.K.A. in-vitro transcription/translation) allows proteins outside of organisms such as E.coli to be recombinantly produced and additionally marked for examination of functional sub-units.

NMR-spectrscopy of biomolecules necessitates high concentrations of the proteins to be tested (generally approx. 1mM in 500µl measured volume) in a very high degree of purity. Molecular biological and biochemical methods are essential for producing such samples.

Another prerequisite for NMR spectroscopy is the existence of nuclei with magnetic quantum number of ½, I.E. 1H, 13C, 15N, 19F and 31P.

Proteins consist of 1H-, 14N- und 12C-Atoms. With the assistance of heterologous or cell-free E.coli expressions we are able to insert the stable isotope 15N (ca. 97%) and 13C (ca. 98%) in proteins and thus render them more accessible for NMR-spectroscopy.

By means of specific marking of (i.e.) single amino acid residues the particularl dynamics and the linked conformation changes of catalytic active proteins can be examined. This “open system” of cell-free synthesis also allows a systematic manipulation of added components so that new knowledge of the interaction of diverse factors of the protein synthesis machinery can be achieved.

Molecules:
P24, CSP, Alzheimer Peptides, AGA2, KanA, Signal transduction proteins,: Ras, Rap, Rho, RalGEF, Byr-kinase and thermophile proteins

Equipment:
Neben der Standardausstattung verfügt unser Labor über
zwei ÄKTA FPLC Chromatographiesysteme und einen Pipettierroboter
MWG Roboseq 4204 SEL Liquid Handling

Contact:
Prof. Dr. Dr. Hans Robert Kalbitzer
Dr. Gudrun Horn

Team:
Emmi Fuchs
Dr. Gerald Bäuml

 

 


Molecular switches
The physiological properties of proteins are mostly determined by their molecular dynamics. Guanine nucleotide binding proteins (GNB-proteins) act as molecular switches in cellular signal transduction, cycling between a GDP-bound „off“-state and a GTP-bound „on“ state. Within our main focus we examine the significance of the molecular dynamics of various GNB-proteins for the on/off switching operation and their effect on the protein-protein interaction of these switch proteins.

For this purpose protein variations from different families of the Ras superfamily, as well as regulatory and signal transduction bonding proteins are recombinantly expressed in E.coli and cleaned with modern FPLC equipment. Various methods of UV/VIS- und fluorescence spectroscopy, high resolution liquid chromatography, Nuclear magnetic resonance spectroscopy (NMR) and in cooperationen micro calorimetry and Electron spin resonance spectroscopy (EPR) are employed for biochemical and biophysical examination.

Contact:
Prof. Dr. Dr. Hans Robert Kalbitzer
Dr. Michael Spörner

Team:
Ina Rosnizeck
Sandra Kreitner
Kerstin Reiß
Maren Schmidt
Tanja Meierhofer
Kathrin Stroka

 

Activation and deactivation of Ras
molecular switches


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Es tut sich was!

30th Trinational Discussion Meeting
Magnetic Resonance in Biology and Chemistry

GDCH Magnetic Resonance Division
Place: Universität Regensburg, Germany
Date: September 22-25, 2008

Freie Stellen für wache Köpfe

Diploma and Phd Theses in Physics, Biology, Chemistry and Bio-Chemistry are always available in our faculty. If you are interested please contact Prof. Kalbitzer and/or team or Secretary,
Tel: 0941 - 943 2595