白念珠菌多药耐药蛋白Cdr1p构效关系研究

发布时间：2018-03-18 04:32

本文选题：白念珠菌　切入点：多药耐药蛋白　出处：《第二军医大学》2006年博士论文　论文类型：学位论文

【摘要】：Cdr1p构效关系的研究有助于深入了解白念珠菌耐药性产生的分子机制。采用定点突变方法获得TMS1(1514L)、TMS2(L563F)、TMS4(W629L和M631I)、TMS6(A772I和L776F)、TMS8(F1239L和M1246Y)和TMS10(A1324L)变异,在酿酒酵母AD1-8u~-菌株中表达,测定表达变异蛋白菌株药物敏感性的改变,罗丹明6G外转运功能的改变,及在氮唑类药物与罗丹明6G共同作用下外转运功能的变化。结果发现L776F、M1246Y和A1324L变异引起菌株对氮唑类药物敏感性明显增加;1514L、L563F、L776F、F1239L、M1246Y和A1324L变异引起菌株外转运罗丹明6G能力的下降;表达野生型蛋白菌株中氟康唑、酮康唑、伊曲康唑和咪康唑与罗丹明6G共同作用时抑制罗丹明6G的外转运,而表达变异蛋白菌株中氮唑类药物与罗丹明6G共同作用下罗丹明6G的外转运能力各异。本研究首次初步证明TMS4、TMS8和TMS10也是Cdr1p重要的底物结合转运位点,发现了与底物结合转运相关的关键性氨基酸残基。
[Abstract]:The study of the structure-activity relationship of Cdr1p is helpful to understand the molecular mechanism of resistance of Candida albicans. By using site-directed mutagenesis method, we obtained the mutagenesis of TMS32011514L, L563FTMS4W629L and M631Ignit6TMI6F1239L and M1246Y), and expressed them in Saccharomyces cerevisiae AD1-8u-, and expressed them in Saccharomyces cerevisiae AD1-8u-. The changes of drug sensitivity and external transport function of Rhodamine 6G were measured. The results showed that the sensitivity of strain L776FU M1246Y and A1324L to azazole increased significantly, and the ability of strain to transport Rhodamine 6G decreased due to the mutation of L776FU M1246Y and A1324L. Fluconazole, ketoconazole, itraconazole and miconazole combined with Rhodamine 6G inhibited the transport of Rhodamine 6G in wild-type protein strains. However, the external transport ability of Rhodamine 6G was different under the interaction of azazole and Rhodamine 6G in the mutant protein expression strain. It was proved for the first time that TMS4TMS 8 and TMS10 were also important substrate-binding transport sites for Cdr1p. Key amino acid residues related to substrate binding transport were found.
【学位授予单位】：第二军医大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R379

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