我们前期发现，能量代谢变化影响白念珠菌对唑类耐药和汉防己甲素逆转耐药的过程。氧化磷酸化是能量代谢的关键环节。为了揭示其机制，本项目针对氧化磷酸化主要环节及其末端酶和关键酶ATP合酶进行研究。拟采用同一亲本来源的氟康唑敏感或耐药菌株，以呼吸链抑制剂、氧化磷酸化抑制剂和解偶联剂分别抑制呼吸链电子传递、ADP磷酸化及其偶联，系统揭示白念珠菌耐药及其逆转与氧化磷酸化各环节的关系。以STA1-Flipper基因敲除和异位高表达术分别构建白念珠菌ATP合酶重要编码基因ATP1、ATP2缺失株和高表达株，观察其用药前后表型、耐药基因表达、蛋白质组学和代谢组学等变化，揭示ATP合酶在耐药及其逆转中的新机制。由于白念珠菌ATP合酶结构特异，项目完成后将极有希望发现氧化磷酸化（尤其ATP合酶）在耐药过程中的新功能，并进而发现作用于该靶点的药物，为抗真菌药物（及其增效剂）的研发提供新思路，具有重要科学意义。

Our previous studies demonstrated that the changes of energy metabolism, in which oxidative phosphorylation is a critical link, are key mechanism underlying azole resistance and tetrandrine-mediated reversal of resistance in Candida albicans. The aim of this project is to reveal the mechanism by investigation of the major steps in oxidative phosphorylation, and ATP synthase, the key enzyme of it. To systematically reveal the the relationship between azole resistance and tetrandrine-mediated reversal of resistance in C. albicans and each step of oxidative phosphorylation, fluconazole-sensitive or fluconazole-resistant substrains of C. albicans isolated from a single patient would be used; the electron transport pathway, phosphorylation of ADP, and Oxidative phosphorylation coupling are suppressed by using of electron transport inhibitors, oxidative phosphorylation inhibitors, and uncoupling agents, respectively. Following this, strains with genes knock-out and overexpression of ATP1 and ATP2, both of which are important coding genes for ATP synthase in C. albicans, would be constructed by STA1-Flipper and overexpression strategies, and observed for differences in phenotype (growth, drug resistance and virulence, etc), proteomics, metabonomics, and other aspects before and after drug administration to discover novel functions as well as mechanisms of ATP synthase in drug resistance and reversible response. Because of the specific structure of ATP synthase in C. albicans, our proposed approach will be great scientific significance for clarifying the novel functions of oxidative phosphorylation in drug resistance, and finding drugs that can act on the specific target, combining anti-fungal therapy and reversing drug resistance.