【作者】 陈宏玲；

【导师】 谢建平；

【作者基本信息】 西南大学 ， 生物化学与分子生物学， 2007， 硕士

【摘要】 结核病，是由结核分枝杆菌引起的疾病，引起全世界每年有300万人死亡。硫在结核杆菌中扮演重要角色。硫代硫酸硫转移酶在细胞硫转运中是一个限速酶，催化转移硫。我们用基因工程方法在大肠杆菌中构建了结核菌硫代硫酸硫转移酶两基因，用硝普钠处理以检测功能。序列比对得到结核菌硫代硫酸硫转移酶中两个保守活性位点(Arg，Cys)，实验中发现加入硝普钠后细胞生存依赖与该酶的表达及硫代硫酸钠有关。探讨一种新的以硝普钠参与的硫代硫酸硫转移酶酶活检测方法和机制。结果预示硫代硫酸硫转移酶在结核菌氰化物解毒和结核铁硫簇形成中可能起作用。而且作为治疗心血管疾病的临床药物硝普钠可能有更广阔的运用。更多还原

【Abstract】 Mycobacterium tuberculosis induces tuberculosis, which causes 3 million deaths annually throughout the world. Sulfur transport plays an important role in Mycobacterium tuberculosis. Thiosulfate sulfurtransferase (TST, EC: 2.8.1.1) is a limiting steps for the sulfur transfer reaction in the cellular, catalyzing the irreversible transfer of sulfane sulfur atom from a suitable donor. We engineered a genetic system to express high levels of recombinant Mycobacterium tuberculosis TST in Escherichia coli, treated the recombinant bacteria under sodium nitroprusside, and used this organism to test the role. Our studies show that conserved residues (Arg, Cys) of TST may be involved in the enzyme activity, and the role of the four thiosulfate sulfiirtransferases of MTB is may similar but not all the same. Under the drug of sodium nitroprusside, recombinant bacteria exhibit endurance. Moreover, we found not only cell viability depend upon expression of TST protein, but also depend upon existence of additive sodium thiosulfate. Then, a new mechanism was described to analyzing the relationship between thiosulfate sulfurtransferase of MTB and sodium nitroprusside. These results indicate that SseB play an important role in cyanide detoxification and Fe—S clusters. Sodium nitroprusside which has been traditionally used to treat high blood pressure may be a universal reagent.更多还原