• Title/Summary/Keyword: platelet microbicidal protein

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Effect of Antibiotics upon the Antibacterial Activity of Platelet Microbicidal Protein against Streptococcus rattus BHT

  • Kim, Jae-Wook;Choe, Son-Jin;Lee, Si-Young
    • International Journal of Oral Biology
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    • v.34 no.1
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    • pp.43-48
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    • 2009
  • Thrombin-induced platelet microbicidal protein (tPMP) is a small cationic peptide that exerts potent in vitro microbicidal activity against a broad spectrum of human pathogens, including Staphylococcus aureus and Streptococcus rattus BHT. Earlier evidence has suggested that tPMP targets and disrupts the bacterial membrane. However, it is not yet clear whether membrane disruption itself is sufficient to kill the bacteria or whether subsequent, presumably intracellular, events are also involved in this process. In this study, we investigated the microbicidal activity of rabbit tPMP toward S. rattus BHT cells in the presence or absence of a pretreatment with antibiotics that differ in their mechanisms of action. The streptocidal effects of tPMP on control cells (no antibiotic pretreatment) were rapid and concentration-dependent. Pretreatment of S. rattus BHT cells with either penicillin or amoxicillin (inhibitors of bacterial cell wall synthesis) significantly enhanced the anti-S. rattus BHT effects of tPMP compared with the effects against the respective control cells over most tPMP concentration ranges tested. On the other hand, pretreatment of S. rattus BHT cells with tetracycline or doxycycline (30S ribosomal subunit inhibitors) significantly decreased the streptocidal effects of tPMP over a wide peptide concentration range. Furthermore, pretreatment with rifampin (an inhibitor of DNA-dependent RNA polymerase) essentially blocked the killing of S. rattus BHT by tPMP at most concentrations compared with the respective control cells. These results suggest that tPMP exerts anti-S. rattus BHT activity through mechanisms involving both the cell membrane and intracellular targets.

Alterations in Cytoplasmic Membrane are Associated with the Bactericidal Activity of Thrombin-Induced Platelet Microbicidal Proteins in Oral Streptococci

  • Choi, Young-Eun;Cheong, Yong-Joon;Lee, Si-Young
    • International Journal of Oral Biology
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    • v.30 no.4
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    • pp.125-130
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    • 2005
  • Thrombin-induced platelet microbicidal proteins (tPMP) are antibacterial proteins released when platelets are stimulated by thrombin. It has been reported that tPMP has antibacterial activity against various bacterial species including causative agents of infective endocarditis. Most of the oral streptococci have resistance to the killing by tPMP and this fact may play an important role as a virulence factor in infective endocarditis. However, the susceptibility and resistance mechanism of oral streptococci for tPMP have not been revealed yet. In this study, the killing mechanism of tPMP for oral streptococci has been investigated. Streptococcus rattus BHT, a susceptible strain, and Streptococcus gordonii DL1, a resistant strain, have been used in this study. tPMP was isolated from platelet after stimulation with thrombin. Cell membrane depolarization was examined with 3,3'-dipropylthiodicarbocyanine iodide ($DiSC_3$), membrane potential-sensitive cyanine dye, by fluorescence spectrophotometry. The permeabilization of cell membrane by tPMP was investigated with propidium iodide (PI) by flow cytometry. tPMP susceptible S. rattus BHT showed the increase of the $DiSC_3$ fluorescence level meaning depolarization of cell membrane and increase of the uptake of PI which means permeabilization of cell membrane. However, tPMP resistant S. gordonii DLI did not show depolarization and permeabilization. These results indicate that the increasing depolarization and permeabilization of oral streptococcal cell membrane are associated with the bactericidal activity of tPMP.