为了探究拟南芥和酿酒酵母来源的磷脂:甘油二酯酰基转移酶(PDAT)在油脂积累过程中与底物相互作用的机制,本研究利用蛋白三维结构模拟、分子对接等生物信息学手段对植物、真菌等来源的PDAT进行蛋白进化关系分析以及PDAT与特定底物的分子对接分析.通过模拟PDAT和磷脂分子的空间对接模型,预测出底物和蛋白空间互作的多个关键氨基酸位点.基于多种磷脂分子和拟南芥PDAT(AtPDAT)的分子对接和氨基酸序列比对,AtPDAT氨基酸序列中的W153,E310,D313,V627可能对蛋白质和底物结合发挥重要作用.该结果精确定位AtPDAT及其它来源的PDAT与磷脂底物的结合位点以及催化位点,为下一步AtPDAT蛋白定向改造提供重要的数据支撑.%To explore the interaction mechanism between substrate and phospholipid: diacylglycerol acyltransferases (PDATs) in Arabidopsis thaliana and Saccharomyces cerevisiae, phylogenetic relationships of PDAT in plants, fungi, and other species were investigated by bioinformatics approaches including three-dimensional structure simulation and molecular docking. The binding model between PDAT and specific phospholipid was created by molecular docking. The key amino acids of intermolecular interactions between phospholipid substrate and PDAT were then predicted. Amino acids including W153, E310, D313, V627 in protein sequences of AtPDAT might have played an important role in substrate-binding domain on the base of molecular docking between several phospholipids of AtPDAT and sequence alignment of PDAT proteins. This result accurately revealed the binding site of substrate to AtPDAT as well as the catalytic domain of candidate PDATs, and provided guidance for future improvement of AtPDAT.
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