Martemyanov K A, Liljas A, Gudkov A T
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, 142292, Russia.
Biochemistry (Mosc). 1998 Oct;63(10):1216-9.
Oligonucleotide-directed mutagenesis was used to obtain elongation factor G from Thermus thermophilus with the G16V mutation in its GTP-binding domain. Functional studies of the mutated protein and elongation factor G from E. coli were carried out. The data revealed that the G16V mutant retains high thermostability, has an increased ribosome-dependent GTPase activity, and its translation activity in cell-free translation system is equal to that of the factor G from E. coli. The mutated protein with an uncleavable GTP analog also has an increased affinity to the ribosomes.
采用寡核苷酸定向诱变技术，从嗜热栖热菌中获得了在其GTP结合结构域具有G16V突变的延伸因子G。对突变蛋白和大肠杆菌延伸因子G进行了功能研究。数据显示，G16V突变体保留了高热稳定性，核糖体依赖性GTP酶活性增加，并且其在无细胞翻译系统中的翻译活性与大肠杆菌的因子G相当。带有不可切割GTP类似物的突变蛋白对核糖体的亲和力也有所增加。
