Viable Coxiella burnetii organisms were isolated from the culture medium of persistently infected Baby Hamster Kidney (BHK-21) fibroblasts. When these organisms were incubated in host-cell-free medium at low pH, some of the de novo-synthesized protein made by the bacteria was translocated to the exterior of the cell. The exported protein was detectable after 2-7 h incubation at 37 degrees C. No evidence was found to suggest that protein accumulation in the medium was due to leakiness caused by cell damage. Both DCCD (dicyclohexylcarbodiimide) and CCCP (carbonyl cyanide m-chlorophenylhydrazone) inhibited the process to some extent. Exported protein was represented largely by three polypeptides with molecular masses of 34, 24 and 12 kDa. De novo-synthesized proteins corresponding to these molecular masses were not detected in cytoplasmic fractions, but a membrane fraction might possess a similar form. It was concluded that a physiological process of protein translocation occurred in C. burnetii during acid activation in a defined medium. Organisms that were extracted directly from the cytoplasm of infected fibroblasts by a mechanical disruption procedure were also active in de novo protein synthesis; however they exported much less of the protein.