Protein capable of binding to the upstream gene of the CYP2B subfamily P450

能够与 CYP2B 亚家族 P450 上游基因结合的蛋白质

• 批准号: 05671829
• 负责人: YAMADA Hideyuki
• 金额: $ 1.34万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671829/
• 关键词: Cytochrome P450; CYP2B subfamily; Barbie box; Phenobarbital; DNA binding protein; Guinea pig; Rat; 核タンパク質; 遺伝子上流; 転写促進

The extent in the induction of hepatic CYP2B P-450 with phenobarbital is greatly different in rats and guinea pigs. To answer this difference the binding of protein to the oligonucleotide probe of-89 to-73 bp region (Barbie box) of CYP2B1 gene was compared in both animal species. When the nuclear extracts of liver were used as the protein fraction, all preparations from phenobarbital-pretreated rats showed strong binding to the Barbie box, but many preparations from untreated rats had only weak activities. On the contrary, the binding with liver nuclear extracts of guinea pigs was not increased by phenobarbital treatment. Therefore, the response to phenobarbital treatment on the nuclear protein-probe interaction was markedly different in rats and guinea pigs. Since this difference correlated with the contrast of the inducibility between rat and guinea pig CYP2B P-450 protein, it was suggested that the expression and induction of the CYP2B P-450 is related with the interaction of nuclea … More r proteins and Barbie box region of the gene. Alteration of the binding between the probe and rat nuclear protein with chemical effectors, and by heating and protease treatments was also examined. The results showed that 1)Ca2+ and Mg2+ effectively increase the binding ; 2)hemin and Triton X-100 but not phenobarbital and dexamethasone increase the binding ; and 3)nuclear protein resists to the heat-denaturation until 60。C and to the proteolysis with proteases. On the other hand, the former two effects were not seen or the extents were limited in the binding with guinea pig liver nuclear extracts. The guinea pig protein capable of binding to the probe were less stable against heating and were much sensitive against proteases than rat protein. The ability of cytosol to bind Barbie box was compared in rats and guinea pigs, and the results are also reported. A guinea pig protein (BBBP-GP) was purified from liver nuclear extracts by monitoring its binding activity toward DNA probe to electrophoretically homogeneous. The minimum molecular weight was estimated to be 27,000. Rat nuclear protein was also purified though only partially. The analysis of the N-terminal sequence indicated that BBBP-GP is a new protein. This protein interacted with oligonucleotide probe, but the shifted band was stretched. Less

在大鼠和豚鼠中，用苯巴比妥诱导肝脏CYP2B P-450的程度大不相同。为了回答这种差异，在两种动物种类中比较了蛋白质与CYP2B1基因的寡核苷酸探针的结合-89 to-73 bp区域（芭比娃娃盒）。当将肝脏的核提取物用作蛋白质分数时，所有来自苯巴比妥预处理大鼠的制剂表现出与芭比娃娃盒的较强结合，但是未经治疗的大鼠的许多制剂只有较弱的活性。相反，苯巴比妥治疗不会增加与豚鼠肝核提取物的结合。因此，在大鼠和豚鼠中，对核蛋白探针相互作用的苯巴比妥治疗的反应明显不同。由于这种差异与大鼠和豚鼠CYP2B P-450蛋白之间诱导性的对比有关，因此建议CYP2B P-450的表达和诱导与核酸核的相互作用有关……基因的更多R蛋白和芭比娃娃盒区域。还检查了通过化学作用以及加热和蛋白质处理的探针和大鼠核蛋白之间结合的改变。结果表明，1）Ca2+和Mg2+有效增加了结合； 2）Hemin和Triton X-100，但没有苯巴比妥和地塞米松增加结合； 3）核蛋白对热饱和的反应物，直到60。c并用蛋白质解析蛋白水解。另一方面，前者没有看到两种作用，或者在与豚鼠肝核提取物结合中的延伸量受到限制。能够与探针结合的豚鼠蛋白对加热的稳定性较低，并且对蛋白质的敏感性比大鼠蛋白敏感得多。在大鼠和豚鼠中比较了细胞质结合芭比娃娃盒的能力，还报告了结果。通过监测其对DNA探针对电泳均匀的探针的结合活性，从肝核提取物中纯化了豚鼠蛋白（BBBP-GP）。最小分子量估计为27,000。大鼠核蛋白也仅部分纯化。 N末端序列的分析表明BBBP-GP是一种新蛋白质。该蛋白质与寡核苷酸探针相互作用，但偏移的带拉伸。较少的