J4 ›› 2012, Vol. 38 ›› Issue (3): 551-554.

• 基础研究 • 上一篇    下一篇

牙龈卟啉单胞菌细胞分裂蛋白质FtsZ的GTPase活性的热稳定性

于维先1|王闻天2|刘歆婵*|张玉凤3   

  1. (1. 吉林大学口腔医院 牙发育及颌骨重塑与再生省重点实验室| |吉林 长春 |130021;2. 吉林大学口腔医院麻醉科|吉林 长春 130021;3. 吉林大学口腔医院修复科|吉林 长春 130021)
  • 收稿日期:2011-12-12 出版日期:2012-05-28 发布日期:2012-05-28
  • 通讯作者: 张玉凤(Tel:0431-88796000,E-mail:yu-wei-xian@163.com) E-mail:yu-wei-xian@163.com
  • 作者简介:于维先(1963-),女,吉林省农安县人,教授,医学博士,主要从事口腔微生态与龋病及牙周病关系的研究。
  • 基金资助:

    吉林省自然科学基金资助课题(201115104)

Thermostability of GTPase activity of cell division protein FtsZ in  Porphyromonas gingivalis

YU Wei-xian1,WANG Wen-tian2,LIU Xin-chan*,ZHANG Yu-feng3   

  1. (1. Laboratory of Mechanism of Tooth Development and Bone Remodeling and Regeration,Jilin Province,Stomatology Hospital,Jilin University,Changchun 130021,China;2. Department of Anesthesiology, Stomatology Hospital,Jilin University,Changchun 130021,China;3. Department of Prosthodontics,Stomatology Hospital,Jilin University,Changchun 130021,China)
  • Received:2011-12-12 Online:2012-05-28 Published:2012-05-28

摘要:

目的:探讨牙龈卟啉单胞菌(Pg)细胞分裂蛋白质FtsZ(PgFtsZ)的GTPase活性的热稳定性,阐明PgFtsZ的生物化学特性,为牙周病的治疗提供实验依据。方法:采用Malachite green assay法检测GTPase的活性,活性测定蛋白质为重组野生型PgFtsZ(WtPgFtsZ)和4种重组C末端缺失变异型PgFtsZ,即ZΔC01(C末端缺失73个氨基酸残基)、ZΔC02(C末端缺失128个氨基酸残基)、ZΔC03(C末端缺失177个氨基酸残基)和ZΔC04(C末端缺失233个氨基酸残基),各种PgFtsZ在未经处理时及煮沸加热处理5和10 min后检测GTPase的酶活力值。结果:重组ZΔC04 GTPase的活性在煮沸加热处理5和10 min时均比未加热处理时的酶活性增加(P<0.05),重组WtPgFtsZ、ZΔC01、ZΔC02和ZΔC03在煮沸加热处理5 min时GTPase的活性均较未煮沸加热处理时的活性增加(P<0.05),但进一步煮沸加热处理10 min时,其GTPase的活性与未加热处理时相似。结论:Pg FtsZ是一种耐热蛋白质,并且C末端缺失233个氨基酸残基的缺失变异型ZΔC04表现出更强的耐热性能。

关键词: 牙龈卟啉单胞菌;重组细胞分裂蛋白质;丝状温度敏感蛋白质;GTPase活性;热稳定性

Abstract:

Abstract:Objective To investigate the thermostability of GTPase activity of cell division protein FtsZ in Porphyromonas gingivalis(Pg) and to discuss the biochemical properties of FtsZ, and to provide experimental basis for curing periodontal disease.Methods Malachite green assay was used to measure the  the GTPase activities of recombinant Wt PgFtsZ,ZΔC01(missing 73 residues from the C-terminus of PgFtsZ),ZΔC02(missing 128 residues from the C-terminus of PgFtsZ),ZΔC03(missing 177 residues from the C-terminus of PgFtsZ),and ZΔC04(missing 233 residues from the C-terminus of PgFtsZ).The GTPase activities of different kinds of proteins were detected  after the protein dealed with unboiling and boiling for 5 and 10 min.The values of GTPase activities were detected at A650nm.Results The GTPase activities of ZΔC04  boiled for 5 and 10 min were higher than those of unboiled(P<0.05).The GTPase activities of WtPgFtsZ,ZΔC01,ZΔC02,and ZΔC03  boiled for 5 min were higher than those of unboiled(P<0.05).But the GTPase activities of WtPgFtsZ,ZΔC01,ZΔC02,and ZΔC03  boiled for 10 min were similar to those of unboiled.Conclusion Pg FtsZ  is a kind of  protein with thermostability,and ZΔC04 missing 233 residues from the C-terminus has higher thermostability.

Key words: Porphyromonas gingivalis, recombinant cell division protein;filamentous temperature sensitive protein Z, GTPase activity, thermostability

中图分类号: 

  • R781.4