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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.
Hepatitis B virus X protein (HBx) plays a pivotal role as a multifunctional regulator of hepatitis-induced liver cancer, with extensive reports of the function of HBx in vitro and the inability of HBx proteins to be solubilized and stabilized in vitro Of the reports of very few.We based on the most common truncated mutant liver cancer patients, we recombinantly expressed in E. coli inclusion bodies retained a 12-127 amino acids, the N-terminal and C-terminal truncated mutant HBxΔNΔC, And renatured to renaturation using renaturation.The molecular sieve showed HBxΔNΔC as a monomer; circular dichroism showed HBxΔNΔC containing 5% α-helix, 30% β-sheet and 65% other structures. HBxΔNΔC can pull down the tumor suppressor p53 and DDB1 proteins from human HepG2 cells and stimulate the migration of liver cells.The intraperitoneal injection of 1 μg / g HBxΔNΔC can significantly stimulate the growth of liver in mice.These results show that HBxΔNΔC has some biological activity, it can be used to study the structure and molecular mechanism of HBx in vitro.