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

苯巴比妥对大鼠和豚鼠肝脏CYP 2BP-450的诱导程度有很大差异。为了回答这一差异，在两种动物中比较了蛋白质与CYP 2B 1基因-89至-73 bp区域（Barbie box）的寡核苷酸探针的结合。当肝的核提取物被用作蛋白质组分时，所有来自苯巴比妥预处理的大鼠的制剂均表现出与芭比盒的强结合，但许多来自未处理的大鼠的制剂仅具有弱活性。与此相反，与豚鼠肝核提取物的结合没有增加苯巴比妥治疗。因此，大鼠和豚鼠对苯巴比妥处理对核蛋白-探针相互作用的反应明显不同。由于这种差异与大鼠和豚鼠CYP 2BP-450蛋白诱导活性的差异有关，提示CYP 2BP-450蛋白的表达和诱导与细胞核蛋白的相互作用有关。 ...更多信息 r蛋白和Barbie box区域。改变探针和大鼠核蛋白与化学效应器之间的结合，并通过加热和蛋白酶处理进行了检查。结果表明：Ca ~（2+）和Mg ~（2+）能有效地促进结合;氯化血红素和Triton X-100能促进结合，而苯巴比妥和地塞米松不能促进结合;核蛋白在60 ℃前能抵抗热变性。C和蛋白酶的蛋白水解。与豚鼠肝细胞核提取物结合时，前两种作用均未观察到或作用程度有限。与大鼠蛋白相比，能够与探针结合的豚鼠蛋白对加热不太稳定，对蛋白酶更敏感。本文比较了大鼠和豚鼠细胞质结合芭比盒的能力。从豚鼠肝细胞核提取物中纯化出一种蛋白质（BBBP-GP），通过检测其与DNA探针的结合活性，使其具有电泳均一性。最小分子量估计为27，000。大鼠核蛋白也被纯化，但只有部分。N端序列分析表明BBBP-GP是一个新蛋白。该蛋白与寡核苷酸探针相互作用，但移动的条带被拉伸。少