Raman microspectroscopy reveals long-term extracellular activity of Chlamydiae.

摘要: The phylum Chlamydiae consists exclusively of obligate intracellular bacteria. Some them are formidable pathogens humans, while others occur as symbionts amoebae. These genetically intractable bacteria possess a developmental cycle consisting replicative reticulate bodies and infectious elementary bodies, which believed to be physiologically inactive. Confocal Raman microspectroscopy was applied differentiate between Protochlamydia amoebophila demonstrate in situ the labelling this amoeba symbiont after addition isotope-labelled phenylalanine. Unexpectedly, uptake amino acid also observed for both stages up 3 weeks, if incubated extracellularly with labelled phenylalanine, P. remained infective during period. Furthermore, energizes its membrane performs protein synthesis outside host. Importantly, extended extracellular incubation could demonstrated human pathogen Chlamydia trachomatis, synthesizes stress-related proteins under these conditions shown by 2-D gel electrophoresis MALDI-TOF/TOF mass spectrometry. findings change our perception chlamydial biology reveal that host-free analyses previously not recognized potential direct experimental access elusive microorganisms.