Protein synthesis or translation is a process critical to life performed by all living organisms. The rapid and accurate production of proteins allows organisms to perform cellular functions, reproduce, and react to environmental stimuli. The core theme of this thesis is prokaryotic translation, beginning with building tools to study the structural dynamics of elongation factor thermo unstable (EF-Tu) and using them to determine the order of events during its functional cycle. Additionally, the phenomenon of prokaryotic structure-based translation initiation by the intergenic region internal ribosome entry site was investigated and its mode of action determined to be different from the previously proposed model. Finally, the preparation and biophysical characterization of the translation initiation region of the rpsA gene from Escherichia coli was performed to begin the work towards an atomic level understanding of this mRNA molecule bound to the prokaryotic ribosome. Together, this thesis demonstrates the robustness of the prokaryotic translation machinery and highlights that, although significant insight has been gained into how these molecular machines operate, there is still additional information needed to fully understand this fundamental process.