Analysis of TNT (2,4,6-trinitrotoluene)-inducible cellular responses and stress shock proteome in Stenotrophomonas sp. OK-5.
In this study, the cellular responses of Stenotrophomonas sp. OK-5 to explosive 2,4,6-trinitrotoluene (TNT) have been extensively analyzed. The stress shock proteins, which might contribute to enhancing cellular resistance to TNT-mediated toxicity, were induced at different concentrations of TNT used as a substrate for cell culture of Stenotrophomonas sp. OK-5 capable of utilizing TNT. Proteomic analysis for 2-DE of soluble protein fractions from the culture of OK-5 exposed to TNT demonstrated approximately 300 spots on the silver-stained gel ranging from pH 3 to pH 10. Among them, 10 spots significantly induced and expressed in response to TNT were selected and analyzed. As the result of internal amino acid sequencing with ESI-Q TOF mass spectrometry, TNT-mediated stress shock proteins such as DnaK, OmpW, and OsmC were identified and characterized. Survival of strain OK-5 was periodically monitored in the presence of different concentrations of TNT along with the production of the stress shock proteins. Cells of strain OK-5 pre-exposed to TNT had in improved survival tolerance. Analysis of total cellular fatty acids in strain OK-5 suggested that several saturated or unsaturated fatty acids might increase or decrease under TNT-mediated stress condition. Scanning electron microscopy of cells treated with 0.8 mM TNT for 12 h revealed irregular rod shapes with wrinkled surfaces.
Department of Life Science, Soonchunhyang University, Asan, P.O. Box 97, Chung-Nam, 336-600, Korea.
Eun-Mi
EM
Ho
Hyo-Won
HW
Chang
Seung-Il
SI
Kim
Hyung-Yeel
HY
Kahng
Kye-Heon
KH
Oh
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001673:Biodegradation, Environmental; D015180:Electrophoresis, Gel, Two-Dimensional; D004785:Environmental Pollutants / Q000378:metabolism / Q000494:pharmacology; D005227:Fatty Acids / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D020543:Proteome / Q000187:drug effects / Q000302:isolation & purification; D020588:Stenotrophomonas / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D014303:Trinitrotoluene / Q000378:metabolism / Q000494:pharmacology*
D001426:Bacterial Proteins; D004785:Environmental Pollutants; D005227:Fatty Acids; D020543:Proteome; D014303:Trinitrotoluene