CAJ | 학술논문

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。
목적：전기연구표명,인TF기인제15호외현자처P589S변이여유양내력상관련。재차기출상,분석해위점변이대TF단백질C2결구역적영향,이탐토기영향유양내력적궤제。방법：응용생물신식학방법,이TF단백C2결구역서렬위기출,용DNAStar Protean연건제공적모괴,예측P589S돌변대기C2결구역적이급결구、단백골가구적유인성、표면가능성、항원계수이급친수성지수적영향。응용분자생물학방법,인공합성함유P589（TF（P））화S589（TF（S））적C2결구역서렬,구건원핵표체중조질립,재E coli.중분별진행표체,원이색보검측병비교불동표체단백이급결구적차이。결과：연건예측결과현시,당제589안기산유P돌변위S시,C2결구역중안기산적이급결구발생료개변,재돌변위점부근,안기산적표면가능성급친수성균하강,이항원계수칙정현출선승고후하강적현상。단백골가구적유인성몰유발생변화。원이색보검측결과현시,량충단백CD보적부봉최저봉도출현재약205 nm처,단TF（P）적봉도요저우TF（S）。계산결과표명,TF（P）단백적α라선비례교TF（S）단백고,기타결구비례상차불대。결론：전철단백P589S다태위점돌변후,단백질C2결구역적이급결구화단백표면성질발생료개변,저충변화장도치기인적상관공능개변,병성위영향유양내력적인소지일。
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.