Award Details:
Award Type: Catalyst
Award #: C-041
Proposal Title: Engineering Prokaryotic Translation for Artificial Metalloenzyme Production
PI(s): Michael Jewett (NU) and Jared Lewis (UChicago)
Award Amount: $200,000.00
Application Cycle: Round 15 (Spring 2013)
Award Start Date: September 1, 2013
Award End Date: August 31, 2015
Abstract:
The extraordinary synthetic capability of protein biosynthesis has driven extensive efforts to engineer translation systems to create proteins containing unnatural amino acids (UAAs). However, UAA incorporation efficiency is ca. 1000x lower than that of natural AAs due to the challenge of re-engineering a system of nearly 80 separate macromolecules. We will identify and eliminate bottlenecks in this system to enable incorporation of organometallic (M)UAAs into proteins. Specifically, we will study MUAA bioavailability, explore interactions between EF-Tu and MUAA-tRNAs, and evolve the entire translation system for maximal MUAA incorporation efficiency. Our work will expand understanding of the translation apparatus, provide a general platform to incorporate any UAA of interest, and facilitate direct expression of a new class of artificial metalloenzymes (ArMs). ArMs combine the selectivity and evolvability of enzymes with the bond forming capabilities of organometallic catalysts. We are particularly interested in non-biogenic metal complexes (e.g., Pd, Ir, and Rh) to promote site-specific C-H functionalization reactions. Such reactions are not currently possible using small molecule catalysts, and they could greatly expand the scope of metabolic engineering for chemical synthesis and enable new methods for direct interrogation of biomolecules in vivo.