Monothiol glutaredoxins can bind linear [Fe3S4]+ and [Fe4S4]2+ clusters in addition to [Fe2S2]2+ clusters: spectroscopic characterization and functional implications.
Monothiol glutaredoxins can bind linear [Fe3S4]+ and [Fe4S4]2+ clusters in addition to [Fe2S2]2+ clusters: spectroscopic characterization and functional implications.
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DOI：
10.1021/ja407059n
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发表时间：
2013-10-09
影响因子：
15
通讯作者：
Johnson MK
中科院分区：
文献类型：
--
作者：
Zhang B;Bandyopadhyay S;Shakamuri P;Naik SG;Huynh BH;Couturier J;Rouhier N;Johnson MK
酿酒酵母线粒体谷氧还蛋白5(Grx5)是一类普遍存在的单一硫醇谷氧还蛋白的典型成员，具有严格保守的CGFS活性位点序列，已被证明在生物[Fe2S2]2+簇运输中起作用。在这项工作中，我们证明了重组酿酒酵母Grx5在无细胞提取物长时间暴露于空气中或在谷胱甘肽存在下进行厌氧重组后进行有氧纯化后，主要包含一个线性的[Fe3S4]+簇。用紫外-可见吸收/CD/MCD、EPR、穆斯堡尔谱和共振拉曼光谱研究了线型[Fe3S4]+团簇的激发态电子性质、基态电子性质和振动性质。结果表明，合成的线状[Fe_3S_4]~+团簇和紫色乌头酶中的线状[Fe_3S_4]~+团簇的性质与文献报道的相似，得到了一个菱形的S=5/2线状[Fe_3S_4]~+团簇。此外，结果表明，以前报道的许多单硫醇GRX的Fe-S团簇含量被完全错误地解释为[Fe2S2]2+团簇，而不是线性[Fe3S4]+团簇或线性[Fe3S4]+和[Fe2S2]2+团簇的混合物。在没有GSH的情况下，Grx5的厌氧重组产生了含有一个[Fe4S4]2+簇的二聚体形式，该簇能够通过完整的簇转移在体外激活脱氧乌头酸酶。通过光谱、突变和分析研究，对Grx5中线型[Fe3S4]+和[Fe4S4]2+团簇的连接进行了研究。根据这些结果，讨论了单硫醇Grx5在清除和回收氧化应激条件下蛋白质展开过程中释放的线状[Fe3S4]+簇以及含有[Fe4S4]2+簇的蛋白质成熟过程中的潜在作用。
Saccharomyces cerevisiae mitochondrial glutaredoxin 5 (Grx5) is the archetypical member of a ubiquitous class of monothiol glutaredoxins with a strictly conserved CGFS active-site sequence that has been shown to function in biological [Fe2S2]2+ cluster trafficking. In this work, we show that recombinant S. cerevisiae Grx5 purified aerobically after prolonged exposure of the cell-free extract to air or after anaerobic reconstitution in the presence of glutathione, predominantly contains a linear [Fe3S4]+ cluster. The excited state electronic properties and ground state electronic and vibrational properties of the linear [Fe3S4]+ cluster have been characterized using UV-visible absorption/CD/MCD, EPR, Mössbauer and resonance Raman spectroscopies. The results reveal a rhombic S = 5/2 linear [Fe3S4]+ cluster with properties similar to those reported for synthetic linear [Fe3S4]+ clusters and the linear [Fe3S4]+ clusters in purple aconitase. Moreover, the results indicate that the Fe-S cluster content previously reported for many monothiol Grxs has been misinterpreted exclusively in terms of [Fe2S2]2+ clusters, rather than linear [Fe3S4]+ clusters or mixtures of linear [Fe3S4]+ and [Fe2S2]2+ clusters. In the absence of GSH, anaerobic reconstitution of Grx5 yields a dimeric form containing one [Fe4S4]2+ cluster that competent for in vitro activation of apo-aconitase, via intact cluster transfer. The ligation of the linear [Fe3S4]+ and [Fe4S4]2+ clusters in Grx5 has been assessed by spectroscopic, mutational and analytical studies. Potential roles for monothiol Grx5 in scavenging and recycling linear [Fe3S4]+ clusters released during protein unfolding under oxidative stress conditions and in maturation of [Fe4S4]2+ cluster-containing proteins are discussed in light of these results.
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影响因子：
15
作者：
GIRERD, JJ;PAPAEFTHYMIOU, GC;HOLM, RH
通讯作者：
HOLM, RH
影响因子：
4.8
作者：
Couturier, Jeremy;Stroeher, Elke;Rouhier, Nicolas
通讯作者：
Rouhier, Nicolas
影响因子：
4.8
作者：
Fernandes, AP;Fladvad, M;Vlamis-Gardikas, A
通讯作者：
Vlamis-Gardikas, A
影响因子：
2.9
作者：
Chandramouli, Kala;Unciuleac, Mihaela-Carmen;Johnson, Michael K.
通讯作者：
Johnson, Michael K.
DOI：
10.1073/pnas.94.12.6087
发表时间：
1997-06-10
影响因子：
11.1
作者：
Khoroshilova, N;Popescu, C;Kiley, PJ
通讯作者：
Kiley, PJ