In vivo metallothionein and glutathione status in an acute response to cadmium in Mercenaria mercenaria brown cells

Gerald Zaroogian, Eugene Jackim
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引用次数: 33

Abstract

Brown cells that are found in the red glands of Mercenaria mercenaria accumulate, detoxify and excrete cadmium. Brown cell involvement in metal detoxification was due in part to endogenous glutathione (GSH) and protein sulfhydryl. Metallothionein (MT) and GSH have been shown to play an important role in metal detoxification in bivalve molluscs. This study showed that the protein sulfhydryl in brown cells of Mercenaria was in fact MT, that brown cell GSH functioned in acute protection against Cd2+ toxicity, that GSH provided the initial defense against Cd2+ toxicity prior to MT induction and that MT variants were unequal in response to Cd2+. During treatment of Mercenaria with 0.5 and 1.0 ppm Cd2+, brown cells were analyzed for MT by capillary electrophoresis and GSH colorimetrically after 0.25, 1, 2, 3, and 4 days. The data indicated that the cadmium-binding protein was MT with an apparent molecular weight of 9 kDa determined by gel filtration or 6 kDa as indicated by capillary electrophoresis. Glutathione appeared to prevail in the brown cell acute response to 0.5 ppm Cd2+, whereas MT appeared to prevail in the acute response to 1.0 ppm Cd2+. Capillary electrophoresis can be used to monitor and quantify MT and its variants in brown cells without need for prior separation of cytosolic components by chromatography. The change in MT-II was greater relative to the change in MT-I in the brown cell acute response to 0.5 ppm Cd2+, whereas the change in MT-I was greater relative to the change in MT-II in the acute response to 1.0 ppm Cd2+. The variants of brown cell MT appeared to respond differentially to Cd2+ depending upon the Cd2+ treatment concentration.

金属硫蛋白和谷胱甘肽在佣兵褐细胞对镉急性反应中的体内状态
在雇佣兵的红色腺体中发现的棕色细胞积聚,解毒和排泄镉。棕色细胞参与金属解毒的部分原因是内源性谷胱甘肽(GSH)和蛋白质巯基。金属硫蛋白(MT)和谷胱甘肽已被证明在双壳类软体动物的金属解毒中起重要作用。本研究表明,佣兵鱼棕色细胞中的巯基蛋白实际上是MT,棕色细胞GSH具有抗Cd2+毒性的急性保护作用,GSH在MT诱导之前提供了抗Cd2+毒性的初始防御,MT变异对Cd2+的反应是不平等的。在0.5和1.0 ppm Cd2+处理下,分别在0.25、1、2、3和4天后,用毛细管电泳和谷胱甘肽比色法分析棕色细胞的MT。结果表明,该镉结合蛋白为MT,凝胶过滤测定其表观分子量为9 kDa,毛细管电泳测定其表观分子量为6 kDa。谷胱甘肽似乎在棕色细胞对0.5 ppm Cd2+的急性反应中占优势,而MT似乎在1.0 ppm Cd2+的急性反应中占优势。毛细管电泳可用于监测和定量MT及其变异在棕色细胞不需要事先分离的细胞质成分的色谱。在0.5 ppm Cd2+的棕色细胞急性反应中,MT-II的变化相对于MT-I的变化更大,而在1.0 ppm Cd2+的急性反应中,MT-I的变化相对于MT-II的变化更大。棕色细胞MT的变体似乎对Cd2+的反应不同,这取决于Cd2+的处理浓度。
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