Reduction of 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt (XTT) is dependent on CaFRE10 ferric reductase for Candida albicans grown in unbuffered media
Reduction of 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt (XTT) is dependent on CaFRE10 ferric reductase for Candida albicans grown in unbuffered media
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DOI：
10.1099/mic.0.28843-0
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发表时间：
2006-08-01
期刊：
影响因子：
2.8
通讯作者：
Dancis, Andrew
中科院分区：
文献类型：
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作者：
Knight, Simon A. B.;Dancis, Andrew
2，3-双（2-甲氧基-4-硝基-5-磺基苯基）-2H-四唑鎓-5-甲酰苯胺内盐（XTT）和其他四唑鎓盐的还原被广泛用作细菌、真菌和哺乳动物细胞活力的测定，但编码还原酶活性的基因尚未被确定。在这里，它表明，XTT和质膜铁还原酶活性是10-40倍，在白色念珠菌比在酿酒酵母。XTT还原酶活性在C.白念珠菌生长在低铁条件下相比，铁充分的条件下，和细胞生长在无缓冲（pH 4（.）0-4（.）4)培养基中，XTT还原酶活性在很大程度上依赖于CaFRE 10。C.白色念珠菌在缓冲至pH 6的培养基中生长（.）8是独立的CaFRE 10，但尽管如此，在细胞中的铁剥夺上调。2-（4，5-二甲基-2-噻唑基）-3，5-二苯基-2H-溴化四唑（MTT），一种膜渗透性四唑盐，发生在细胞内的位置，是独立的CaFRE 10。而C. albicans，而S.啤酒。C.白色念珠菌具有多种铁和pH值调节还原酶的活动，能够减少四唑盐，但在无缓冲介质中生长时，CaFRE 10所需的XTT还原酶活性。
The reduction of 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt (XTT) and other tetrazolium salts is widely used as an assay for bacterial, fungal and mammalian cell viability, but the genes encoding the reductase activities have not been defined. Here, it was shown that XTT and plasma membrane ferric reductase activities were 10-40-fold greater in Candida albicans than in Saccharomyces cerevisiae. XTT reductase activity was induced fivefold in C. albicans grown in low-iron conditions compared with iron-replete conditions, and for cells grown in unbuffered (pH 4(.)0-4(.)4) medium, XTT reductase activity was largely dependent on CaFRE10. XTT reductase activity of C. albicans grown in medium buffered to pH 6(.)8 was independent of CaFRE10 but, nonetheless, was upregulated in cells deprived of iron. Reduction of 2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide (MTT), a membrane-permeable tetrazolium salt, occurred at an intracellular location and was independent of CaFRE10. However, MTT activity was induced by iron deprivation in C. albicans but not in S. cerevisiae. C. albicans possessed multiple iron- and pH-regulated reductase activities capable of reducing tetrazolium salts, but, when grown in unbuffered medium, CaFRE10 was required for XTT reductase activity.