The Mitochondria-to-Cytosol H(2)O(2) Gradient Is Caused by Peroxiredoxin-Dependent Cytosolic Scavenging.
The Mitochondria-to-Cytosol H(2)O(2) Gradient Is Caused by Peroxiredoxin-Dependent Cytosolic Scavenging.
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线粒体到胞质溶解醇H（2）O（2）梯度是由过氧蛋白依赖性的胞质清除引起的。
DOI：
10.3390/antiox10050731
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发表时间：
2021-05-06
期刊：
影响因子：
--
通讯作者：
Hidalgo E
中科院分区：
文献类型：
--
作者：
de Cubas L;Pak VV;Belousov VV;Ayté J;Hidalgo E
开发了用于测量细胞内H2 O2的基于荧光蛋白的报告物，以克服小的可渗透染料的局限性。遗传编码的氧化还原报告基因的两个主要家族是融合至过氧化物还原蛋白和Hyperer及其衍生物的基于还原-氧化敏感性绿色荧光蛋白（roGFP）的蛋白质。我们使用了每个家族中最敏感的探针roGFP 2-Tpx1.C169S和Hyper 7来监测裂变酵母中过氧化物的稳态和波动水平。虽然两者都能够监测细胞内H2 O2的纳摩尔波动，但前者的灵敏度是HyPer 7的2 - 5倍，并且roGFP 2-Tpx1.C169S在野生型细胞的胞质溶胶中被部分氧化，而HyPer 7被完全还原。我们已经成功地在线粒体基质中表达了Hyper 7，并且它被氧化了约40%，这表明在低微摩尔范围内，过氧化物的稳态水平高于胞质溶胶。除非不存在主要的H2 O2清除剂（细胞质过氧化物氧还蛋白Tpx 1），否则细胞质HyPer 7可以检测到线粒体来源的细胞质中可忽略不计的H2 O2，而线粒体HyPer 7在野生型和Δ tpx 1细胞中的氧化程度相同。我们的结论是，有一个双向流量的H2 O2跨矩阵和胞质溶胶，但Tpx 1迅速有效地清除代谢产生的过氧化物，并停止其稳态胞质水平上升。
Fluorescent protein-based reporters used to measure intracellular H2O2 were developed to overcome the limitations of small permeable dyes. The two major families of genetically encoded redox reporters are the reduction-oxidation sensitive green fluorescent protein (roGFP)-based proteins fused to peroxiredoxins and HyPer and derivatives. We have used the most sensitive probes of each family, roGFP2-Tpx1.C169S and HyPer7, to monitor steady-state and fluctuating levels of peroxides in fission yeast. While both are able to monitor the nanomolar fluctuations of intracellular H2O2, the former is two-five times more sensitive than HyPer7, and roGFP2-Tpx1.C169S is partially oxidized in the cytosol of wild-type cells while HyPer7 is fully reduced. We have successfully expressed HyPer7 in the mitochondrial matrix, and it is ~40% oxidized, suggesting higher steady-state levels of peroxides, in the low micromolar range, than in the cytosol. Cytosolic HyPer7 can detect negligible H2O2 in the cytosol from mitochondrial origin unless the main H2O2 scavenger, the cytosolic peroxiredoxin Tpx1, is absent, while mitochondrial HyPer7 is oxidized to the same extent in wild-type and ∆tpx1 cells. We conclude that there is a bidirectional flux of H2O2 across the matrix and the cytosol, but Tpx1 rapidly and efficiently scavenges mitochondrial-generated peroxides and stops their steady-state cytosolic levels rising.
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DOI：
10.1042/bj20081386
发表时间：
2009-01-01
期刊：
The Biochemical journal
影响因子：
--
作者：
Murphy MP
通讯作者：
Murphy MP
影响因子：
4.1
作者：
Han, D;Williams, E;Cadenas, E
通讯作者：
Cadenas, E
影响因子：
14.8
作者：
Morgan, Bruce;Van Laer, Koen;Dick, Tobias P.
通讯作者：
Dick, Tobias P.
影响因子：
11.4
作者：
Antunes F;Brito PM
通讯作者：
Brito PM
影响因子：
3.6
作者：
Paulo, Esther;Garcia-Santamarina, Sarela;Hidalgo, Elena
通讯作者：
Hidalgo, Elena