Effect of the metal ion-induced carbonylation modification of mitochondrial membrane channel protein VDAC on cell vitality, seedling growth and seed aging.
Effect of the metal ion-induced carbonylation modification of mitochondrial membrane channel protein VDAC on cell vitality, seedling growth and seed aging.
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DOI：
10.3389/fpls.2023.1138781
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发表时间：
2023
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
--
作者：
种子是种质资源保存的最重要载体。然而，种子成熟后，活力可能会发生不可逆转的下降，这称为种子老化。线粒体是种子老化过程中启动细胞程序性死亡的重要细胞器。然而，潜在的机制仍不清楚。我们前期的蛋白质组研究发现，在榆树种子老化过程中，有13种线粒体蛋白发生了羰基化修饰。本研究通过固定化金属亲和层析(IMAC)检测到金属结合蛋白，表明线粒体中的金属结合蛋白是种子老化过程中碳化的主要目标。采用生物化学、分子和细胞生物学方法检测金属-蛋白质结合、蛋白质修饰和亚细胞定位。利用酵母和拟南芥对其体内生物学功能进行了研究。在IMAC实验中，包括线粒体电压依赖性阴离子通道(VDAC)在内的12个蛋白被鉴定为Fe2+/Cu2+/Zn2+结合蛋白。UpVDAC与上述三种金属离子均有结合能力。His204Ala(H204A)和H219A突变的UpVDAC蛋白失去了与金属的结合能力，对金属催化氧化(MCO)诱导的羰化反应不敏感。野生型UpVDAC的过表达使酵母细胞对氧化胁迫更加敏感，抑制了拟南芥幼苗的生长，加速了种子的衰老，而突变的UpVDAC的过表达减弱了VDAC的这些作用。这些结果揭示了金属结合能力与羰基化修饰的关系，以及VDAC可能在调节细胞活力、幼苗生长和种子衰老方面的作用。
Seeds are the most important carrier of germplasm preservation. However, an irreversible decrease in vigor can occur after the maturation of seeds, denoted as seed aging. Mitochondrion is a crucial organelle in initiation programmed cell death during seed aging. However, the underlying mechanism remains unclear. Our previous proteome study found that 13 mitochondria proteins underwent carbonylation modification during the aging of Ulmus pumila L. (Up) seeds. This study detected metal binding proteins through immobilized metal affinity chromatography (IMAC), indicating that metal binding proteins in mitochondria are the main targets of carbonization during seed aging. Biochemistry, molecular and cellular biology methods were adopted to detect metal-protein binding, protein modification and subcellular localization. Yeast and Arabidopsis were used to investigate the biological functions in vivo. In IMAC assay, 12 proteins were identified as Fe2+/Cu2+/Zn2+ binding proteins, including mitochondrial voltage dependent anion channels (VDAC). UpVDAC showed binding abilities to all the three metal ions. His204Ala (H204A) and H219A mutated UpVDAC proteins lost their metal binding ability, and became insensitive to metal-catalyzed oxidation (MCO) induced carbonylation. The overexpression of wild-type UpVDAC made yeast cells more sensitive to oxidative stress, retarded the growth of Arabidopsis seedlings and accelerated the seed aging, while overexpression of mutated UpVDAC weakened these effects of VDAC. These results reveal the relationship between the metal binding ability and carbonylation modification, as well as the probable function of VDAC in regulating cell vitality, seedling growth and seed aging.
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影响因子：
3.7
作者：
Maisonneuve E;Ducret A;Khoueiry P;Lignon S;Longhi S;Talla E;Dukan S
通讯作者：
Dukan S
影响因子：
4.3
作者：
Colombini, M
通讯作者：
Colombini, M
影响因子：
56.9
作者：
Hiller, Sebastian;Garces, Robert G.;Wagner, Gerhard
通讯作者：
Wagner, Gerhard
影响因子：
7.4
作者：
Salvato, Fernanda;Havelund, Jesper F.;Moller, Ian Max
通讯作者：
Moller, Ian Max
影响因子：
3.4
作者：
Kung, Cheng-Che S.;Huang, Wei-Ning;Yeh, Kuo-Chen
通讯作者：
Yeh, Kuo-Chen