Using genetic code expansion to investigate the functional dynamics of proteins

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Proteins are the key functional parts of the cell. Their dynamic 3D-structure determines their activity. It is controlled by stimuli like the binding of ligands or post-translational modifications. In order to understand how cells function we must elucidate the dynamic behaviour of proteins. Conformational changes in proteins can be detected by introducing biophysical probes, however, this is often technically very difficult. Here, I suggest a strategy to overcome this limitation that introduces unnatural amino acids (UAAs) with unique biophysical properties using components for genetic code expansion (evolved orthogonal ribosomes and tRNA/aminoacyl-tRNA-synthetase pairs), which I have recently created. This will allow to incorporate multiple different UAAs into the same protein and therefore to introduce FRET pairs, which can be used to decipher the dynamics of proteins and its implication for their molecular mechanism of function. I intend to exploit this unique technology to investigate the impact of histone core acetylations, which can only be obtained in pure form by genetically encoding acetyllysine on recombinant histones and have been inaccessible to biochemical analysis so far, on the dynamics of chromatin assembly and remodelling.