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BP: Fachverband Biologische Physik

BP 28: Protein structure and dynamics II

BP 28.7: Talk

Wednesday, April 2, 2014, 16:45–17:00, HÜL 386

Genetically Encoded Spin Labelled Artificial Amino Acids — •Malte Drescher, Moritz Schmidt, and Daniel Summerer — Konstanz Research School Chemical Biology and Department of Chemistry, University of Konstanz, Germany

Recent publications demonstrate the ability of electron paramagnetic resonance spectroscopy (EPR) to provide structural, dynamical and functional data on biomacromolecules in cells. Of particular interest are distance measurements in the nanometer range. The advantages of the method are sensitivity, selectivity, the lack of any limitation imposed by the size of the macromolecule, and the possibility to get information on coexisting conformations via analyzing distance distributions. However, so far, these approaches require microinjection of spin-labelled macromolecules. Moreover, the biomolecules transferred to cells by these means have limited access to natural mechanisms of cellular processing like folding, localization, posttranslational modification and natural decay.

Here, we show for the first time the successfull incorporation of a genetically encoded modified lysine amino acid containing 2, 2, 5, 5, -tetramethyl-pyrrolin-1-oxyl-label into various positions in GFP and TRX mutants in E.coli. First EPR distance measurements on extracted proteins demonstrate the potential of this novel approach.