Recent years have seen the convergence of both genetic and biochemical approaches in the study of protein translocation in E. coli. The powerful combination of these approaches is exemplified in the use of an in vitro protein synthesis-protein translocation system to analyze the role of genetically defined components of the protein translocation machinery. We describe in this review recent results focusing on the function of the secA, secB, and secY gene products and the demonstration of their requirement for in vitro protein translocation. The SecA protein was recently shown to possess ATPase activity and was proposed to be a component of the translocation ATPase. We present a speculative working model whereby the translocator complex is composed of the integral membrane proteins SecY, SecD, SecE, and SecF, forming an aqueous channel in the cytoplasmic membrane, and the tightly associated peripheral membrane protein SecA functioning as the catalytic subunit of the translocator or "protein-ATPase."