天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (7): 705-709.doi: 10.11958/j.issn.0253-9896.2015.07.001

• 细胞与分子生物学 •    下一篇

酸性条件下变异链球菌临床株表面相关蛋白表达差异分析

赵兴福， 江山，黄晓晶，闫福华   

  1. 1天津市口腔医院牙体牙髓二科 （邮编 300041）； 2福建医科大学附属口腔医院牙体牙髓科； 3南京大学医学院附属口腔医院
  • 收稿日期:2014-12-16 修回日期:2015-03-08 出版日期:2015-07-15 发布日期:2015-07-15
  • 通讯作者: 黄晓晶 E-mail:hxiaoj@163.com E-mail:xingfu_108@126.com
  • 作者简介:赵兴福 （1981）， 男， 硕士， 主治医师， 主要从事龋病病因学研究
  • 基金资助:
    国家自然科学基金资助项目 （30500564）

Differential expressions of surface-associated proteins of streptococcus mutans strains isolated from clinical samples under acid condition

ZAHO Xingfu, JIANG Shan, HUANG Xiaojing, YAN Fuhua   

  1. 1 Department of Endodontology, Tianjin Stomatological Hospital of Nankai University, Tianjin 300041, China; 2 Department of Endodontics and Operative Dentistry, School and Hospital of Stomatology, Fujian Medical University; 3 Institute and Hospital of Stomatology, Nanjing University Medical School
  • Received:2014-12-16 Revised:2015-03-08 Published:2015-07-15 Online:2015-07-15
  • Contact: HUANG Xiaojing E-mail:hxiaoj@163.com E-mail:xingfu_108@126.com

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摘要： 目的 探讨变异链球菌临床株在 pH7.0 和 pH5.0 条件下的表面相关蛋白表达差异， 进一步了解龋病发生和发展过程。方法 采用 Homer 法提取 593 和 18 号 （593 号菌株分离于高发龋患者， 18 号菌株分离自无龋健康人）临床分离株在 pH7.0 和 pH5.0 条件下的表面相关蛋白， 经十二烷基硫酸钠聚丙烯酰胺凝胶电泳和二维电泳确定蛋白表达的差异条带及位点， 差异位点由基质辅助激光解析电离飞行时间质谱分析， 并结合数据库进行检索和鉴定蛋白质。结果 经过对电泳图谱的分析发现， 两菌株在 pH7.0 条件下存在 4 个高表达位点和 2 个特异位点； 在 pH5.0 条件下存在 2 个高表达位点和 2 个特异位点。593 号菌株蛋白表达又发生了特殊的变化， 即在 pH5.0 条件下， 存在 3个高表达位点和 6 个特异位点， 其中双组分信号传导系统组氨酸激酶 Lyts 和 3-磷酸甘油醛脱氢酶高表达， NADH 氧化酶特异表达。结论 两菌株在酸性变化下均出现了某些蛋白的高表达和特异表达， 可能两菌株耐酸反应中存在相似之处； 两菌株间蛋白表达差异可能是耐酸性存在差异的原因。

关键词: 链球菌, 变异, 蛋白质组学, 临床分离株, 表面相关蛋白, 耐酸性

Abstract: Objective To investigate the changes in the surface-associated protein expression of streptococcus rnutans (Sm) isolated from clinical samples at pH7.0 and pH5.0. Methods The proteins were extracted from cells at pH7.0 and pH5.0 by Homer method. The proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE) and two-dimensional gel electrophoresis followed by image analysis. Proteins were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry and computer-assisted protein sequence analysis. Results Image analysis revealed that four high expression levels of protein loci and two specific protein loci were existed in two strains at pH7.0. Two high expression levels of protein loci and two specific protein loci were existed in two strains at pH5.0. Three high expression levels of protein loci and six specific protein loci were existed in Sm593 at pH5.0. Two-component system histidine kinases Lyts and glyceraldehyde- 3- phosphate dehydrogenase were highly modulated, and NADH oxidase were modulated specifically. Conclusion The two clinical isolations in acid have high expression of some special proteins, which are presumed to be the resemblance of acid reaction．The difference of protein expression of two clinical isolations in acid may represent their distinct acid resistance．

Key words: streptococcus mutans, proteomics, the clinical isolationns, the surface-associated proteins, acid resistance