Improved tag-switch method reveals that thioredoxin acts as depersulfidase and controls the intracellular levels of protein persulfidation.
Improved tag-switch method reveals that thioredoxin acts as depersulfidase and controls the intracellular levels of protein persulfidation.
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DOI：
10.1039/c5sc04818d
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发表时间：
2016-05-25
期刊：
影响因子：
8.4
通讯作者：
Filipovic MR
中科院分区：
文献类型：
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作者：
Wedmann R;Onderka C;Wei S;Szijártó IA;Miljkovic JL;Mitrovic A;Lange M;Savitsky S;Yadav PK;Torregrossa R;Harrer EG;Harrer T;Ishii I;Gollasch M;Wood ME;Galardon E;Xian M;Whiteman M;Banerjee R;Filipovic MR
H2S信号通过蛋白质过硫化。要成为监管机构，这种修改必须是可逆的。使用一种新的过硫化物检测方法，我们发现了这个缺失的环节，并表明硫氧还蛋白系统在体内起着脱过硫化物酶的作用。硫化氢（H2S）已经成为一种信号分子，能够调节几种重要的生理功能，如血压，神经传递和炎症。这些影响背后的机制仍然是很大程度上难以捉摸的半胱氨酸残基的氧化翻译后修饰（蛋白质过硫化或S-硫化）已被提出作为硫化氢诱导的生物学和药理学效应的主要途径。作为一种信号机制，必须控制过硫化。使用改进的标签开关检测过硫化物，我们在这里表明，蛋白质过硫化物水平控制的硫氧还蛋白系统。重组硫氧还蛋白对过硫化半胱氨酸的反应性几乎是对胱氨酸的10倍，并且易于裂解蛋白质过硫化物。该反应导致H2S释放，表明硫氧还蛋白可能是过硫化物池中H2S水平的重要调节剂。硫氧还蛋白系统的抑制引起细胞内过硫化物的增加，突出硫氧还蛋白作为控制H2S信号传导的主要蛋白脱过硫化物酶。最后，使用来自具有较高循环硫氧还蛋白水平的HIV-1患者的血浆，我们可以证明脱过硫酶在体内的作用。
H2S signals via protein persulfidation. To be regulatory the modification will have to be reversible. Using a new method for persulfide detection, we discover this missing link and show that thioredoxin system acts as depersulfidase in vivo. Hydrogen sulfide (H2S) has emerged as a signalling molecule capable of regulating several important physiological functions such as blood pressure, neurotransmission and inflammation. The mechanisms behind these effects are still largely elusive and oxidative posttranslational modification of cysteine residues (protein persulfidation or S-sulfhydration) has been proposed as the main pathway for H2S-induced biological and pharmacological effects. As a signalling mechanism, persulfidation has to be controlled. Using an improved tag-switch assay for persulfide detection we show here that protein persulfide levels are controlled by the thioredoxin system. Recombinant thioredoxin showed an almost 10-fold higher reactivity towards cysteine persulfide than towards cystine and readily cleaved protein persulfides as well. This reaction resulted in H2S release suggesting that thioredoxin could be an important regulator of H2S levels from persulfide pools. Inhibition of the thioredoxin system caused an increase in intracellular persulfides, highlighting thioredoxin as a major protein depersulfidase that controls H2S signalling. Finally, using plasma from HIV-1 patients that have higher circulatory levels of thioredoxin, we could prove depersulfidase role in vivo.