摘要
甾醇转运蛋白在促进植物生长发育、提高逆境胁迫耐受性、降低血浆低密度脂蛋白以及胆固醇等作用中发挥重要作用.杜仲(Eucommia ulmoides)是一种甾醇含量丰富且具有调节血压等多种功用的名贵中药材,目前对杜仲甾醇转运蛋白NPC1 及其编码基因的分子信息还不清楚.本研究根据人类NPC1 基因信息进行杜仲EuNPC1-Like基因的发掘和分析鉴定.从杜仲基因组中获得的EuNPC1-Like基因全长3876 bp,编码蛋白属于不稳定疏水性脂溶蛋白;分别含有20 个糖基化位点和136 个磷酸化位点,是由13 个α螺旋组成的跨膜蛋白,主要分布于溶酶体/液泡和细胞膜,具有NPC1_N胆固醇转运保守结构域,其蛋白三维结构与人类和拟南芥的NPC1 相似,含有光、赤霉素、低温、干旱、厌氧、茉莉酸甲酯响应等顺式作用元件,与向日葵、油橄榄、咖啡和樟子松等油脂含量高的植物NPC1 蛋白亲缘关系较近.网络药理学分析表明,NPC1 蛋白参与植物甾醇代谢调控,杜仲β-谷甾醇靶向人体脂肪与固醇代谢,与动植物中的甾醇代谢相似.本研究结果为揭示EuNPC1-Like基因功能提供了依据.
Abstract
Sterol transporters are known to have a significant impact on various aspects of plant physiology,including growth and development,stress tolerance,and the regulation of plasma low density lipoprotein and cholesterol levels in the human body.Eucommia ulmoides,a Chinese medicinal material renowned for its sterol content,has various health benefits such as regulating blood pressure.However,little is known about the molecular characteristics of the sterol transporter NPC1 and its corresponding gene in E.ulmoides.In this study,the EuNPC1-Like gene of Eucommia ulmoides was identified,analyzed,and characterized using knowledge from the human NPC1 gene.The results showed that the EuNPC1-Like protein had a total length of 3876 base pairs and was characterized as an unstable hydrophobic lipid soluble protein.This protein possessed 20 glycosylation sites and 136 phosphorylation sites.It was a transmembrane protein consisting of 13 α helices,primarily localized in lysosomes/vacuoles and cell membranes.Furthermore,it contained a conserved cholesterol transport domain known as NPC 1_N,which exhibited a three-dimensional structure similar to that of NPC1 protein in humans and Arabidopsis.Additionally,the protein contains cis-acting elements that responded to various stimuli such as light,gibberellin,low temperature,drought,anaerobic conditions,and methyl jasmonate.The NPC1 protein was closely associated with high oil content plants such as sunflower,olive,coffee,and pinus camphoris.Network pharmacological analysis suggested that the NPC 1 protein played a role in regulating phytosterol metabolism.Additionally,Eucommia β-sitosterol targeted human fat and sterol metabolism,which was similar to sterol metabolism in plants and animals.This study provides a basis for further understanding the function of the EuNPC1-Like gene.
基金项目
贵州省科技计划(黔科合服企2018-4001-06)
贵州省科技计划(黔科合基础ZK-2023-119)
贵州省研究生科研基金(黔教合2020-079)