Proteome analysis of cellular response of Pseudomonas putida KT2440 to tetracycline stress.
Tetracycline-induced proteome of Pseudomonas putida KT2440 was analyzed by 2-D gel electrophoresis and matrix-assisted laser desorption ionization-time of flight/mass spectrum (NALDI-TOF/MS) in order to understand cellular response to tetracycline. Of the proteins upregulated in a culture medium containing subinhibitory concentration of tetracycline (50 mug/mL), we identified 38 proteins from cytosol and precipitated fractions by peptide mass fingerprinting and mass spectrum/mass spectrum analysis. Various amino acids ABC transporters, a ribose ABC transporter, and a sulfate ABC transporter were found to be upregulated. Protein synthesis-related proteins, stress proteins, energy metabolic enzymes, and unknown proteins were also strongly induced. Of the identified upregulated proteins, several proteins (isocitrate lyase, branched-chain amino acid ABC transporter, superoxide dismutase, etc.) were also upregulated under phenol-induced stress condition. These results demonstrate that tetracycline at a high concentration induced comprehensive stress in P. putida KT2440 and the global induction of proteins related to bacteria survival. Proteome analysis was found to be a useful tool for the elucidation of antibiotic-induced proteins in the present study.
Yun
Sung-Ho
SH
Proteomics Team, Korea Basic Science Institute, Daejeon, 305-333, Korea.
Kim
Young Hwan
YH
Joo
Eun Jin
EJ
Choi
Jong-Soon
JS
Sohn
Jung-Hoon
JH
Kim
Seung Il
SI
D001426:Bacterial Proteins / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D020543:Proteome; D016958:Pseudomonas putida / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013752:Tetracycline / Q000494:pharmacology*; D013753:Tetracycline Resistance / Q000235:genetics; D015854:Up-Regulation
D001426:Bacterial Proteins; D020543:Proteome; D013752:Tetracycline