(6cq) Engineering the Ribosome for the Development of New Technologies

Contreras, L. M., Cornell University

Engineering the Ribosome for the Development of New Technologies Recent data has linked ribosomes to a number of important cellular processes that extend beyond protein biosynthesis; some of these include protein folding, secretion regulation and targeting, and translation regulation. The realization that ribosomes dynamically connect polypeptide synthesis to other major cellular processes has led to the development of several powerful biotechnological applications such as a method known as in vitro ribosome display for isolating improved functional protein variants from large combinatorial libraries. Additionally, several key properties of ribosomes have led to the possibility of extending its use for protein purification and solubility enhancement. While the development of these technologies has been facilitated by the vast amount of structural and dynamic data that has been collected on the ribosome in recent years, a great deal about this complex machinery remains unknown. In particular, lack of knowledge about how the interactions that take place during protein translocation inside the ribosome may impact the downstream processing of proteins has hindered the development of new technologies and the extension of already existing ones. Our research has focused on exploring how the ribosome can be further used to improve the soluble expression of complex proteins in bacteria by: (1) understanding how the passage of proteins from the ribosome to the cytosol can affect its folding properties, (2) developing in vivo ribosome-mediated technologies for applications in protein engineering based on the interactive properties of the ribosome, and (3) engineering the ribosome for an efficient protein purification tool. Our progress so far indicates that ribosomes are remarkably ?engineerable? and hold great promise for the development of numerous diverse technologies.