【摘 要】
：
由结核分枝杆菌(Mycobacterium tuberculosis)引起的结核病严重危害人类的健康.结核分枝杆菌具有特殊的细胞壁结构,与细菌增殖和存活、感染、致病性、免疫逃逸等有关.细胞壁核心结构由分枝菌酸、聚阿拉伯半乳糖和肽聚糖构成,分枝菌酸-聚阿拉伯半乳糖通过二糖衔接分子：L-鼠李糖-D-N-乙酰葡糖胺与肽聚糖共价连接.在细胞壁中,还存在非共价结合的糖脂、细胞壁蛋白,其中一些细胞壁蛋白是甘露
【机 构】
：
大连医科大学基础医学院生物化学教研室,大连,116044
【出 处】
：
中国生物化学与分子生物学会2016年全国学术会议
论文部分内容阅读
由结核分枝杆菌(Mycobacterium tuberculosis)引起的结核病严重危害人类的健康.结核分枝杆菌具有特殊的细胞壁结构,与细菌增殖和存活、感染、致病性、免疫逃逸等有关.细胞壁核心结构由分枝菌酸、聚阿拉伯半乳糖和肽聚糖构成,分枝菌酸-聚阿拉伯半乳糖通过二糖衔接分子：L-鼠李糖-D-N-乙酰葡糖胺与肽聚糖共价连接.在细胞壁中,还存在非共价结合的糖脂、细胞壁蛋白,其中一些细胞壁蛋白是甘露糖基化蛋白.分枝杆菌蛋白质O-甘露糖基化过程包括糖基化的起始和糖链的延长.起始过程是由聚戊烯醇-磷酸-甘露糖-蛋白质-甘露糖基转移酶(PMT)将聚戊烯醇-磷酸-甘露糖(PPM)的甘露糖基转移到多肽链中Ser或Thr 的羟基上.延长过程由甘露糖基转移酶催化,将GDP-甘露糖的甘露糖通过α((1,2糖)苷键连接到已有的甘露糖上.结核分枝杆菌(M.tuberculosis)Rv1002c 基因编码PMT,且为细菌生长必需基因.耻垢分枝杆菌(M.smegmatis)是研究结核分枝杆菌的模式菌,其MSMEG_5447 为Rv1002c 的同源基因.在本研究中,我们用同源DNA 重组技术构建了MSMEG_5447 基因敲除菌株(ΔM5447),并确定MSMEG_5447 为细菌生长的非必需基因.ΔM5447 菌株可用于研究结核分枝杆菌O-甘露糖基化蛋白及其在感染宿主细胞中的作用.我们克隆了结核分枝杆菌 Rv1096 基因,构建了分枝杆菌表达载体,分别电转化耻垢分枝杆菌mc2155 野生型菌株和ΔM5447 菌株,获得mc2155/Rv1096 和ΔM5447/Rv1096 菌株.用凝集素ConA 对Rv1096 蛋白进行Western blot 分析,结果显示在mc2155/Rv1096 菌株中表达的Rv1096蛋白发生O-甘露糖基化,而在ΔM5447/Rv1096 菌株中表达的Rv1096 蛋白则不发生O-甘露糖基化.我们将用mc2155/Rv1096 菌株和ΔM5447/Rv1096 菌株分别感染小鼠,进一步研究Rv1096 蛋白的甘露糖链在致病中的作用.
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