转铁蛋白P589S变异影响有氧耐力的机制研究  

Study the Mechanism of P589S Variation in Transferrin Effects on Aerobic Endurance

作　　者：夏小慧[1] 王冬梅[2] 张一英[1] 鄂彤[1] 胡扬[3] 

机构地区：[1]兰州城市学院,甘肃兰州730070 [2]西北民族大学,甘肃兰州730030 [3]北京体育大学,北京100084

出　　处：《北京体育大学学报》2013年第6期65-68,共4页Journal of Beijing Sport University

基　　金：国家自然科学基金(31071032)

摘　　要：目的:前期研究表明,人TF基因第15号外显子处P589S变异与有氧耐力相关联。在此基础上,分析该位点变异对TF蛋白质C2结构域的影响,以探讨其影响有氧耐力的机制。方法:应用生物信息学方法,以TF蛋白C2结构域序列为基础,用DNAStar Protean软件提供的模块,预测P589S突变对其C2结构域的二级结构、蛋白骨架区的柔韧性、表面可能性、抗原系数以及亲水性指数的影响。应用分子生物学方法,人工合成含有P589(TF(P))和S589(TF(S))的C2结构域序列,构建原核表达重组质粒,在E coli.中分别进行表达,圆二色谱检测并比较不同表达蛋白二级结构的差异。结果:软件预测结果显示,当第589氨基酸由P突变为S时,C2结构域中氨基酸的二级结构发生了改变,在突变位点附近,氨基酸的表面可能性及亲水性均下降,而抗原系数则呈现出先升高后下降的现象。蛋白骨架区的柔韧性没有发生变化。圆二色谱检测结果显示,两种蛋白CD谱的负峰最低峰都出现在约205 nm处,但TF(P)的峰度要低于TF(S)。计算结果表明,TF(P)蛋白的α螺旋比例较TF(S)蛋白高,其他结构比例相差不大。结论:转铁蛋白P589S多态位点突变后,蛋白质C2结构域的二级结构和蛋白表面性质发生了改变,这种变化将导致基因的相关功能改变,并成为影响有氧耐力的因素之一。Objective： Previous study shows that the P589S variation in fifteenth exon of transferrin（TF） gene is related to aerobic endurance.The influence of P589S on C2 domain of TF protein was researched to reveal the mechanism of this variation effecting on aerobic endurance.Methods： Using bioinformatic approaches,based on the C2 domain sequences of TF protein,DNAStar software was used to predict the influence of P589S variation on the secondary structure,flexible regions,surface probability plot,antigenic index,and hydrophilicity plot of C2 domain of TF protein,respectively.Using molecular biological approaches,C2 domains coding sequence containing P589 and S589 were syntheticed and the prokaryotic recombinant plasmid were constructed and then expressed in E coli.Circular dichroism spectra（CD） were used to detect and contrast the difference in secondary structure between the two proteins.Results： Software predicted that when 589P converts to 589S,the secondary structure of C2 domain is changed.Close to the mutational site,the surface probability plot and hydrophilicity plot is decreased,and the antigenic index is first increased and then decreased.The flexible regions remain unchanged.Both of the proteins have a CD absorption in 205 nm,but the value of TF（P） is lower than TF（S）.The calculation results showed that the proportion of α-helix is more in TF（P） than that in TF（S）.Conclusions： The variation P589S in TF gene changes the secondary structure of C2 domain and the ability of protein surface.This maybe the mechanism of this variation effects on aerobic endurance.

关 键 词：铁转运蛋白 基因多态性 二级结构 圆二色谱 

分 类 号：G824.2[文化科学—体育训练]