Proteomics-based analysis of a counter-oxidative stress system in Porphyromonas gingivalis.
Porphyromonas gingivalis is a Gram-negative anaerobic pathogen associated with chronic periodontitis. Although anaerobic, P. gingivalis exhibits a high degree of aerotolerance, which enables it to survive within periodontal pockets. The aim of the present study was to examine the effect of oxidative stress on protein expression in P. gingivalis to obtain a better understanding of the mechanism underlying its aerotolerance. To accomplish this, P. gingivalis cells were grown under conditions of hemin limitation (0.01 microg/mL) to avoid the oxygen protective effect of hemin on oxidative stress. The proteins were then extracted from cultures either left untreated or subjected to oxidative stress and separated by 2-DE. The resultant protein expression profiles were examined by image scanning, and those found to differ depending on the presence or absence of aeration were subjected to MALDI-MS and then analyzed using the ORF database of P. gingivalis W83 from The Institute of Genomic Research. Oxidative stress was found to affect the expression of numerous proteins in P. gingivalis cells. In particular, the levels of HtpG, GroEL, DnaK, AhpC, TPR domain protein, and trigger factor were substantially increased.
Okano
Soichiro
S
Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry at Matsudo, 2-870-1 Sakaecho-nishi, Matsudo, Chiba 271-8587 Chiba, Japan.
Shibata
Yasuko
Y
Shiroza
Teruaki
T
Abiko
Yoshimitsu
Y
D001426:Bacterial Proteins / Q000737:chemistry*; D001483:Base Sequence; D017931:DNA Primers; D015180:Electrophoresis, Gel, Two-Dimensional; D016366:Open Reading Frames; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D016966:Porphyromonas gingivalis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D040901:Proteomics; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D001426:Bacterial Proteins; D017931:DNA Primers; D012329:RNA, Bacterial; D012333:RNA, Messenger; D010100:Oxygen