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胶质细胞源性神经营养因子 (GDNF)在神经系统损伤修复中具有重要作用。根据大鼠GDNF晶体结构结果 ,用PCR方法改造人GDNF编码基因 ,在大肠杆菌中表达并纯化获得了一系列人GDNF片段缺失及插入突变体 ,通过对脊髓神经元存活率的测定来观察结构改造对人GDNF神经营养活性的影响。结果表明 ,GDNF分子内部的“胱氨酸结”结构对于GDNF分子构象的维持十分重要 ,GDNF分子中α螺旋、指状结构 1区、指状结构 2区对GDNF神经营养活性的维持必不可少 ,而GDNF的N末端氨基酸残基对其生物活性的维持作用不大。
Glial cell line-derived neurotrophic factor (GDNF) plays an important role in the repair of nervous system injury. According to the results of rat GDNF crystal structure, a series of deletion and insertion mutants of human GDNF fragment were obtained by transforming the human GDNF coding gene by PCR and expressing and purifying in E. coli. The structure alteration was observed by measuring the survival rate of the neurons in the spinal cord Effect on Human Neurotrophic Activity of Human GDNF. The results showed that the “cystine knot” structure of GDNF molecule is very important for maintaining the conformation of GDNF molecule. In GDNF molecule, α helix, finger 1, and finger 2 are necessary for the maintenance of GDNF neurotrophic activity , While the N-terminal amino acid residues of GDNF have little effect on their biological activity.