Crystallization of Escherichia coli 70S Ribosome in complex with tRNAs

Abstract

The prokaryotic ribosome is responsible for translation, the process of creating polypeptides from amino acids. Specifically, ribosomes translate the codons in mRNAs into polypeptide chains. The prokaryotic ribosome is made up of a large subunit and a small subunit, which are known as the 50S (Svedberg units) and 30S subunits, respectively. The larger subunit catalyzes peptide bond formation within the peptidyl transferase center, while the smaller subunit mediates decoding of the mRNA. Together they make up the 70S prokaryotic ribosome. Studies have shown that certain nascent polypeptides can bind to the exit tunnel, resulting in inhibition of peptide bond formation which is referred to as ribosome stalling. Through the use of X‐Ray crystallography we will crystallize the 70S ribosome in complex with tRNAs and a nascent chain within the exit tunnel producing a structure. Although, no structure has been produced yet, we have been able to narrow down conditions that will optimize the growth of these crystals. This optimization will allow for production of the structure which will result in us being able to further explore the process of nascent chain mediated‐translational arrest in the Escherichia coli (E. coli) 70S ribosome.This research was supported in part by a grant from the Howard Hughes Medical Institute.