复性的乙型肝炎病毒X蛋白突变体HBxΔNΔC具有体外生物学活性（英文）

发布时间：2018-11-09 21:35

【摘要】：乙型肝炎病毒X蛋白(HBx)在肝炎引起的肝癌病变中作为一个多功能调节因子起着关键的作用.对HBx的功能有非常多的体外研究报导,而由于HBx蛋白难以溶解和稳定使得体外的研究报导非常少.我们根据肝癌患者中最常见的截短突变体,在大肠杆菌包涵体中重组表达了一个保留了第12~127位氨基酸的、N端和C端截断的突变体HBxΔNΔC,并使用变复性的方法将其复性溶解.分子筛显示HBxΔNΔC为单体;圆二色谱显示HBxΔNΔC包含有5%的α-螺旋,30%的β-折叠和65%的其他结构.复性的HBxΔNΔC能从人HepG2细胞中拉下(pull-down)肿瘤抑制因子p53和DDB1蛋白,它还能刺激肝细胞的迁移.腹腔注射1μg/g的HBxΔNΔC能明显刺激小鼠肝的生长.这些结果显示HBxΔNΔC具有一定的生物学活性,它可以用来研究体外的HBx结构和分子作用机制.
[Abstract]:Hepatitis B virus X protein (HBx) plays a key role as a multifunctional regulatory factor in Hepatitis induced liver cancer. There are many studies on the function of HBx in vitro, but there are very few in vitro due to the difficulty of dissolving and stabilizing the HBx protein. According to the most common truncated mutants in patients with hepatocellular carcinoma, we expressed a mutant HBx 螖 N 螖 C, which retains the amino acid at position 121127in E. coli inclusion body, and refolded and dissolved it by renaturation. The molecular sieve showed HBx 螖 N 螖 C as monomer, and the circular dichroism showed that HBx 螖 N 螖 C contained 5% 伪 -helix, 30% 尾 -folding and 65% other structures. Renatured HBx 螖 N 螖 C can pull down the tumor suppressor (pull-down) p53 and DDB1 proteins from human HepG2 cells, and it can also stimulate the migration of hepatocytes. Intraperitoneal injection of 1 渭 g / g HBx 螖 N 螖 C significantly stimulated liver growth in mice. These results indicate that HBx 螖 N 螖 C has certain biological activity, which can be used to study the structure and molecular mechanism of HBx in vitro.
【作者单位】：复旦大学生命科学学院生理学与生物物理学系;复旦大学生殖与发育研究所;上海工业菌株工程技术研究中心;
【基金】：supported by grants from Chinese High-tech Research and Development Program(No.2013AA102803B,2014AA021301) Shanghai Science and Technology Commission(No.13DZ2252000) Shanghai Leading Academic Discipline Project(Project number B111) National Talent Training Fund in Basic Research of China
【分类号】：R373.21

【相似文献】