Wang Zhongshan, Zhang Meng, Shi Xiaodong, Xiang Quanju
Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical University, Xuzhou, China.
Department of Gynecology, Central Hospital of Xuzhou, Affiliated Hospital of Southeast University, Xuzhou, China.
Biomed Res Int. 2017;2017:3076091. doi: 10.1155/2017/3076091. Epub 2017 Jun 12.
The coding sequence of was cloned and expressed in . The protein was purified and ATPase activity was characterized by NADH oxidation method. GsiA exhibited optimum activity at 30°C and at pH 8 in Tris/HCl buffer. GsiA protein was stable at 20°C. 66% and 44% activity remained after incubation at 30°C and 40°C for 30 min. pH 7 and pH 9 incubation would obviously reduce the ATPase activity. In vivo functionality of was determined by constructing gene deletion strains. was shown to be essential for GSI mediated glutathione uptake and deletion could decrease the virulence of . Interactions of glutathione import proteins GsiA, GsiB, GsiC, and GsiD were investigated by using bacterial two-hybrid system. GsiA could interact with itself and inner membrane proteins GsiC and GsiD. This report provides the first description of functions in . The results could help elucidating the glutathione uptake mechanism and glutathione functions in bacteria.
的编码序列被克隆并在 中表达。该蛋白质被纯化，并且通过NADH氧化法对ATP酶活性进行了表征。GsiA在30°C以及Tris/HCl缓冲液pH 8的条件下表现出最佳活性。GsiA蛋白在20°C下稳定。在30°C和40°C孵育30分钟后，分别保留了66%和44%的活性。在pH 7和pH 9条件下孵育会明显降低ATP酶活性。通过构建基因缺失菌株来确定 的体内功能。结果表明 对于GSI介导的谷胱甘肽摄取至关重要，缺失 会降低 的毒力。通过细菌双杂交系统研究了谷胱甘肽转运蛋白GsiA、GsiB、GsiC和GsiD之间的相互作用。GsiA可以与自身以及内膜蛋白GsiC和GsiD相互作用。本报告首次描述了 在 中的功能。这些结果有助于阐明细菌中谷胱甘肽的摄取机制和谷胱甘肽的功能。
