REGULATION OF ZINC PROTEINS AND CELLULAR ZINC HOMEOSTASIS

锌蛋白和细胞锌稳态的调节

• 批准号: 07457541
• 负责人: OGURI Kazuta
• 金额: $ 0.96万
• 依托单位: FACULTY OF PHARMACEUTICAL SCIENCES,KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457541/
• 关键词: cytosol; aldolase; rat; alcohol dehydrogenase; triose phosphate; tyrosine phosphatase; carbonic anhydrase; polychlorinated biphenyl; ホメオスタシス; 亜鉛フィンガー; 転写調節因子

Toxic manifestation of coplanar PCB is thought to be mediated by aromatic hydrocarbon (Ah)-receptor. However, the suppression of protein expression by coplanar PCBs is scarcely clarified. We report here the suppression of cytosolic proteins by a coplanar PCB-treatment and propose new aspects of the toxicity. Male Wistar rats were treated with PenCB (single 25 mg/kg, i. p.). Free- and pair-fed control groups were treated with vehicle. At the day 5, the liver cytosol was prepared. The cytosolic proteins were compared among three groups by SDS-PAGE and two-dimensional RAGE (2D-PAGE). Expression level of cytosolic 40-kDa and 27-kDa proteins in rat liver were markedly suppressed by PenCB-treatment. The 40-kDa proteins were identified as aldolase B (Ald B) and alcohol dehydrogenase (ADH) class I by amino acid sequencing of the internal peptides. The 27-kDa protein was also identified as carbonic anhydrase III (CA II). The liver cytosolic Ald and ADH activities were significantly reduced to about 50% and 60% of both control groups by PenCB-treatment. Immunoblotting after sD-PAGE demonstrated that Ald B was markedly suppressed by PenCB-treatment, while change in Ald A was slight. The reduction of ADH by PenCB-treatment was also verified by immunoblotting. The significant suppression of CA III by PenCB-treatment in rat liver was demonstrated by immunoblotting using CA III-selective antibody. Ald plays an important role in glycolytic and gluconeogenetic pathways. In addition, ADH catalyzes biotransformation of triose phosphate to glycerol-3-phosphate. Sippression of Ald B and ADH may be a key biochemical lesion for disordered intermediary metabolism occurring by PenCB-treatment and should be taken into an account as a cause of the wasting syndrom which is a severe toxic effect of toxic coplanar PCBs. CA III possesses tyrosine phosphatase activity, therefore, its suppression could account for the effects on signal transduction by toxic coplanar PCBs.

共面印刷电路板的毒性表现被认为是由芳香烃受体介导的。然而，共平面多氯联苯对蛋白质表达的抑制作用几乎还不清楚。我们在这里报告了通过共面多氯联苯处理对胞浆蛋白的抑制，并提出了毒性的新方面。雄性Wistar大鼠灌胃PenCB(单次25 mg/kg，ip)。自由饲养组和配对饲养组均给予赋形剂处理。第5天，制备肝细胞胞浆。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法和双向凝胶电泳法比较三组间胞浆蛋白的差异。大鼠肝细胞胞浆40-kDa和27-kDa蛋白的表达水平明显受到抑制。经氨基酸序列分析，确定该40 kDa蛋白为醛缩酶B(Ald B)和乙醇脱氢酶(ADH)I类。这个27 kDa的蛋白也被鉴定为碳酸氢酶III(CA II)。经PenCB处理后，小鼠肝细胞胞浆Ald和ADH活性明显降低，分别为对照组的50%和60%。SD-PAGE免疫印迹结果显示，经PenCB处理后，Ald B受到明显抑制，而Ald A变化不大。免疫印迹实验也证实了PenCB对ADH的抑制作用。用CA III选择性抗体进行免疫印迹实验表明，PenCB对大鼠肝脏CA III有明显的抑制作用。ALD在糖酵解和糖异生途径中起着重要作用。此外，ADH还催化磷酸三糖生物转化为3-磷酸甘油。AldB和ADH的表达下调可能是经PTCB治疗后中间代谢紊乱的关键生化损害，应作为毒性共面多氯联苯的严重毒性反应--耗竭综合征的原因之一加以考虑。CA III具有酪氨酸磷酸酶活性，因此，它的抑制可能是有毒共面多氯联苯对信号转导的影响。

项目成果

Yuji Ishii et al.: "Significant suppression of rat liver aldolase B by a toxic coplanar polychlorinated biphenyl,3,3',4,4',5-pentachlorobiphenyl" Toxicology. 116. 193-199 (1997)

Yuji Ishii 等人：“有毒的共面多氯联苯,3,3,4,4,5-五氯联苯对大鼠肝脏醛缩酶 B 的显着抑制”毒理学。

Yuji Ishii et al.: "Significant suppression of rat liver aldolase B by a toxic coplanar polychlorinated biphenyl,3,3′,4,4′,5-pentachlorobiohenyl" Toxicology. 116. 193-199 (1997)

Yuji Ishii 等人：“有毒的共面多氯联苯，3,3,4,4,5-五氯二苯基对大鼠肝脏醛缩酶 B 的显着抑制”毒理学。 116. 193-199 (1997)

Ishii, Y., Kato, H., Hatsumura, M., Ishida, T.Ariyoshi, N., Oguri, K.: "Significant suppression of rat liver aldolase B by a toxic coplanar polychlorinated biphenyl, 3,3', 4,4', 5-pentachlorobiphenyl" Toxicology. 116. 193-199 (1997)

Ishii, Y.、Kato, H.、Hatsumura, M.、Ishida, T.Ariyoshi, N.、Oguri, K.：“有毒共面多氯联苯对大鼠肝脏醛缩酶 B 的显着抑制，3,3, 4