Institute of Microbial Technology (Council for Scientific and Industrial Research), Chandigarh 160036, India.
Institut für Molekulare Infektionsbiologie, Universität Würzburg, 97080 Würzburg, Germany.
J Biol Chem. 2011 Dec 2;286(48):41183-41194. doi: 10.1074/jbc.M111.272377. Epub 2011 Oct 12.
Candida albicans lacks the ability to survive within its mammalian host in the absence of endogenous glutathione biosynthesis. To examine the ability of this yeast to utilize exogenous glutathione, we exploited the organic sulfur auxotrophy of C. albicans met15Δ strains. We observed that glutathione is utilized efficiently by the alternative pathway of glutathione degradation (DUG pathway). The major oligopeptide transporters OPT1-OPT5 of C. albicans that were most similar to the known yeast glutathione transporters were not found to contribute to glutathione transport to any significant extent. A genomic library approach to identify the glutathione transporter of C. albicans yielded OPT7 as the primary glutathione transporter. Biochemical studies on OPT7 using radiolabeled GSH uptake revealed a K(m) of 205 μm, indicating that it was a high affinity glutathione transporter. OPT7 is unusual in several aspects. It is the most remote member to known yeast glutathione transporters, lacks the two highly conserved cysteines in the family that are known to be crucial in trafficking, and also has the ability to take up tripeptides. The transporter was regulated by sulfur sources in the medium. OPT7 orthologues were prevalent among many pathogenic yeasts and fungi and formed a distinct cluster quite remote from the Saccharomyces cerevisiae HGT1 glutathione transporter cluster. In vivo experiments using a systemic model of candidiasis failed to detect expression of OPT7 in vivo, and strains disrupted either in the degradation (dug3Δ) or transport (opt7Δ) of glutathione failed to show a defect in virulence.
白色念珠菌在缺乏内源性谷胱甘肽生物合成的情况下,无法在哺乳动物宿主中生存。为了研究这种酵母利用外源性谷胱甘肽的能力,我们利用了白色念珠菌 met15Δ 菌株的有机硫营养缺陷型。我们观察到,谷胱甘肽可被谷胱甘肽降解的替代途径(DUG 途径)有效地利用。白色念珠菌的主要寡肽转运蛋白 OPT1-OPT5 与已知的酵母谷胱甘肽转运蛋白最为相似,但它们对谷胱甘肽的转运没有显著贡献。利用基因组文库方法鉴定白色念珠菌的谷胱甘肽转运蛋白,发现 OPT7 是主要的谷胱甘肽转运蛋白。使用放射性标记的 GSH 摄取对 OPT7 的生化研究表明,K(m)为 205 μm,表明它是一种高亲和力的谷胱甘肽转运蛋白。OPT7 在几个方面都很不寻常。它是与已知的酵母谷胱甘肽转运蛋白最远的成员,缺乏家族中两个高度保守的半胱氨酸,这些半胱氨酸在运输中至关重要,并且还具有摄取三肽的能力。转运蛋白受培养基中硫源的调节。OPT7 同源物在许多致病性酵母和真菌中普遍存在,并形成一个与酿酒酵母 HGT1 谷胱甘肽转运蛋白簇截然不同的独特簇。在系统性念珠菌病模型中的体内实验未能检测到 OPT7 在体内的表达,并且谷胱甘肽降解(dug3Δ)或转运(opt7Δ)缺陷的菌株在毒力方面没有表现出缺陷。