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Date:
2016-10-05
[Abstract] Copper is an essential micronutrient and functions as a cofactor in many enzymes such as heme-Cu oxygen reductases, Cu-Zn superoxide dismutases, multi-copper oxidases and tyrosinases. However, due to its chemical reactivity, free copper is highly toxic (Rae et al., 1999) and all organisms use sophisticated machineries for controlling uptake, storage and export of Cu. The strict control of the cellular Cu homeostasis prevents toxic effects but sustains synthesis of cuproproteins. Monitoring the copper levels within the cell and within different cellular compartments is an essential approach for identifying the contribution of different proteins in maintaining the cellular copper equilibrium. Therefore, whole cells and whole-cell lysates, which can be further fractionated into ...
[摘要] 铜是必需的微量营养素并且在许多酶如辅酶Cu氧还原酶,Cu-Zn超氧化物歧化酶,多铜氧化酶和酪氨酸酶中起辅因子的作用。然而,由于其化学反应性,游离铜是高毒性的(Rae等人,1999),并且所有生物体使用复杂的机器来控制Cu的摄取,储存和输出。严格控制细胞铜稳态防止毒性作用,但维持铜蛋白的合成。监测细胞内和不同细胞区室内的铜水平是识别不同蛋白质在维持细胞铜平衡中的贡献的必要方法。因此,可以进一步分离成细胞质和周质的全细胞和全细胞裂解物被消化,并通过Lowry测定法测定蛋白质浓度。随后,通过原子吸收光谱(AAS)测量铜含量,并计算每mg蛋白质的Cu含量。这提供了产生关于细胞Cu含量的可定量结果的简单且成本有效的方法。为了举例说明这种方法,我们使用光养型α-变形杆菌荚膜红细菌(Rhodobacter capsulatus),其通常用作研究细菌细胞中Cu运输的模式生物体(Ekici等人)。 ,2012)。 [背景] 由于对细胞Cu稳态的兴趣日益增长,在过去几年中开发了用于测量细胞Cu含量的不同方法。它们包括电化学和荧光方案,电感耦合等离子体质谱(ICP-MS),电感耦合等离子体原子发射光谱(ICP-AES),电子微探针分析(EMPA),X射线吸收光谱(XAS)或同步辐射X ...
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