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pH stress가 Streptococcus mutans의 형질막 유동성 및 atpB 유전자 발현에 미치는 영향

pH Stress Alters Cytoplasmic Membrane Fluidity and atpB Gene Expression in Streptococcus mutans

  • 조철민 (부산대학교 치의학전문대학원 구강생화학교실) ;
  • 정승일 (부산대학교 치의학전문대학원 구강생화학교실) ;
  • 김명섭 (부산대학교 치의학전문대학원 구강생화학교실) ;
  • 이새아 (부산대학교 치의학전문대학원 구강생화학교실) ;
  • 강정숙 (부산대학교 치의학전문대학원 구강생화학교실)
  • Cho, Chul Min (Department of Oral Biochemistry and Molecular Biology, School of Dentistry, Pusan National University) ;
  • Jung, Seung Il (Department of Oral Biochemistry and Molecular Biology, School of Dentistry, Pusan National University) ;
  • Kim, Myung Sup (Department of Oral Biochemistry and Molecular Biology, School of Dentistry, Pusan National University) ;
  • Lee, Sae A (Department of Oral Biochemistry and Molecular Biology, School of Dentistry, Pusan National University) ;
  • Kang, Jung Sook (Department of Oral Biochemistry and Molecular Biology, School of Dentistry, Pusan National University)
  • 투고 : 2016.10.25
  • 심사 : 2016.12.22
  • 발행 : 2017.01.30

초록

치아우식의 주원인균인 Streptococcus mutans (S. mutans)는 산 생성 뿐 아니라 산에 대한 탁월한 저항성을 나타낸다. 본 연구에서는 S. mutans가 pH stress에 노출될 때 형질막 유동성, F-ATPase 활성과 발현 및 양성자 투과성 변화와 그 상관관계를 규명하였다. S. mutans로부터 형질막을 분리한 후 1,6-diphenyl-1,3,5-hexatriene을 사용하여 pH stress가 형질막 유동성 변화에 미치는 영향을 측정하였다. pH 4.8과 pH 8.8에서 배양한 S. mutans는 pH 6.8에서 배양한 S. mutans에 비하여 형질막 유동성이 감소되었다. F-ATPase 활성과 발현은 pH 4.8에서 가장 높았고, pH 8.8에서 가장 낮았다. 양성자 투과성은 pH 4.8과 pH 8.8에서 모두 감소되었으며, 특히 pH 4.8에서의 감소가 컸다. F-ATPase 활성만으로 양성자 투과성이 결정된다면 pH 8.8에서 가장 높아야 하나 pH 6.8보다 감소하는 것은 형질막 유동성 감소에 기인된 양성자 세포내 유입 감소와 관련된 것으로 추정한다. 또한 pH 4.8에서 양성자 투과성이 아주 낮은 것은 높은 F-ATPase 활성에 의한 양성자 세포외 유출 증가 뿐 아니라 형질막 유동성 감소에 의한 양성자 세포내 유입 감소에 기인된 것으로 추정한다. 따라서 pH stress에 의한 형질막 유동성 감소는 S. mutans가 세포내 pH 를 유지하는데 중요한 역할을 하는 것으로 생각되며 에탄올을 포함하여 비특이적으로 세포막 유동성을 증가시키는 약물들은 항우식제에 활용될 수 있을 것으로 추정한다.

Streptococcus mutans (S. mutans), which plays a major role in the etiology of human dental caries, is able to tolerate exposure to acid shock in addition to its acidogenicity. We investigated the effects of pH stress on membrane fluidity, activities and expression levels of F-ATPase, and proton permeability in S. mutans. Using 1,6-diphenyl-1,3,5-hexatriene, we observed membrane ordering at pH 4.8 and pH 8.8. The ordering effects were larger at pH 4.8 in cytoplasmic membranes isolated from S. mutans (CMSM). Increasing pH resulted in a decrease in the activities and expression levels of F-ATPase. The proton permeability was decreased at both acidic and alkaline pHs, and the lowest permeability was observed at pH 4.8. The lower permeability at pH 8.8 than pH 6.8 is likely to be caused by the decreased proton influx due to the decreased CMSM fluidity. In addition, it seems to be evident that extremely low permeability at pH 4.8 was caused by the decreased proton influx due to the decreased CMSM fluidity as well as the increased proton efflux due to the increased activity and expression level of F-ATPase. It is likely that CMSM fluidity and F-ATPase activity are two major key factors that determine proton permeability in S. mutans. We suggest that CMSM fluidity plays an important role in the determination of proton permeability, which sheds light on the possibility of using nonspecific membrane fluidizers, e.g., ethanol, for anti-caries purposes.

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피인용 문헌

  1. vol.42, pp.4, 2018, https://doi.org/10.11149/jkaoh.2018.42.4.224