Rap1GAP二聚体的形成对其在细胞内定位的影响  

Influence of Formation of Rap1GAP Dimer on Intracellular Localization of Rap1GAP

作　　者：赵春艳[1] 连培文[1] 魏巍[1] 孔繁斗[2] 

机构地区：[1]大连医科大学检验医学院临床生化教研室,大连116027 [2]大连医科大学附属第一医院妇产科,大连116011

出　　处：《中国生物制品学杂志》2007年第3期195-197,共3页Chinese Journal of Biologicals

摘　　要：目的观察Rap1GAP二聚体的形成对其在细胞内定位的影响。方法对绿色荧光蛋白标记的Rap1GAP野生体编码第90和99位氨基酸的核苷酸进行定点突变,制备绿色荧光蛋白标记的Rap1GAP二聚体缺失突变体M90和M99。将Rap1GAP野生体及二聚体缺失突变体单独或者分别与活化型Gα共转染HEK293细胞,在荧光显微镜下观察野生体和二聚体缺失突变体在细胞内的分布情况。结果野生型Rap1GAP与二聚体缺失突变体Rap1GAP分别单独转染HEK293细胞时,野生型和突变体Rap1GAP均弥漫分布胞质内;当野生型与突变体Rap1GAP分别与活化型Gα共转染HEK293细胞时,野生型和突变体Rap1GAP均主要分布在细胞膜上。结论Rap1GAP二聚体的形成对其在细胞内的定位没有影响。Objective To observe the influence of formation of RaplGAP dimer on the intracellular localization of RaplGAP. Methods Induce the site-directed mutations of nucleotides encoding the amino acids at sites 90 and 99 of green fluorescent protein-labeled wild RaplGAP to obtain mutants Mg0 and M99 with RaplGAP dimer deleted respectively. HEK293 cells were transfected with wild Rap1GAP,mutant M90,mutant M99 ,wild RaplGAP ＋ Get,mutant M90 ＋ Get and M99 ＋ Get respectively. The distributions of wild Rap1GAP and mutants were observed by fluorescent microscopy. Results When HEK293 cells were transfected with wild RaplGAP, mutant M90 or mutant M99,both wild RaplGAP and mutants were diffused in cytoplasm. However,when the cells were co-transfected with wild Rap1GAP ＋ Gα, M90 ＋ Gα or M99 ＋ Get,both wild RaplGAP and mutants were mainly distributed on cell membrane. Conclusion The formation of RaplGAP dimer showed no significant influence on the intracellular localization of Rap1GAP.

关 键 词：Rap1GAP 二聚体 点突变 细胞内定位 

分 类 号：R735.9[医药卫生—肿瘤]