灵芝细胞中磷脂酸互作蛋白鉴定
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金 (Nos. 31900027,31772374,32071673),中国博士后科学基金 (No. 2020M682601),湖南省自然科学基金 (No. 2020JJ5972),湖南省科技创新计划 (No. 2020RC2059),湖南省教育厅科学研究项目 (No. 18B167),中南林业科技大学研究生科技创新基金(No.CX20202014) 资助。


Identification of phosphatidic acid interacting proteins in Ganoderma lingzhi
Author:
Affiliation:

Fund Project:

National Natural Science Foundation of China (Nos. 31900027, 31772374, 32071673), China Postdoctoral Science Foundation (No. 2020M682601), Natural Science Foundation of Hunan Province, China (No. 2020JJ5972), Science and Technology Innovation Program of Hunan Province, China (No. 2020RC2059), Scientific Research Fund of Hunan Provincial Education Department, China (No. 18B167), Scientific Innovation Fund for Post-graduates of Central South University of Forestry and Technology, China (No. CX20202014).

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    灵芝是名贵药用真菌,三萜是灵芝的关键药效成分。前期研究发现,磷脂酶D (Phospholipase D,PLD) 产生的磷脂酸 (Phosphatidic acid,PA) 可调控三萜合成,为进一步阐明PA调控灵芝三萜合成的分子机制,研究采用PA-beads富集结合LC-MS/MS技术,鉴定灵芝细胞中PA互作蛋白,结果共鉴定到了19个PA互作蛋白,主要包括细胞色素P450单加氧酶 (GL22084)、特异性蛋白激酶MAPK (GL23765)、过氧化氢酶和细胞表面疏水性蛋白等。通过基因克隆、原核表达载体构建、蛋白诱导表达和分离纯化,获得了融合GST标签的GL22084和GL23765蛋白,采用GST-pull down实验,验证了灵芝GL22084和GL23765蛋白与PA互作。研究结果揭示了灵芝细胞中PA互作蛋白,为后续解析PLD介导的PA信号分子调控灵芝三萜合成的分子机理奠定了基础;同时,鉴定到的PA互作蛋白也为其他物种的PLD/PA信号通路相关研究提供借鉴。

    Abstract:

    Ganoderma lingzhi is widely recognized as a medicinal basidiomycetes. Triterpene acids (TAs) are the key bioactive medicinal components of G. lingzhi. Our previous studies have shown that phospholipid acid (PA) produced by phospholipase D (PLD) plays a regulatory role in TA synthesis. In order to further elucidate the molecular mechanism how PA regulates TA synthesis in G. lingzhi, PA beads enrichment combined with LC-MS/MS technology was used to identify PA interacting proteins in G. lingzhi. A total of 19 PA interacting proteins were identified, including cytochrome P450 monooxygenase (GL22084), specific protein kinase MAPK (GL23765), catalase and cell surface hydrophobicity-associated protein. GST tagged GL22084 and GL23765 proteins were obtained through gene cloning, heterologous expression, and purification. The interactions between GL22084/GL23765 and PA were verified by GST pull down assay. The identification of PA interacting proteins provides a basis for further understanding the molecular mechanism how PLD-mediated PA signaling molecules regulates the TA synthesis in G. lingzhi. Moreover, the PA interacting proteins identified in this study can also provide clues for the research of PLD/PA signaling pathway in other species.

    参考文献
    相似文献
    引证文献
引用本文

刘勇男,尹媛媛,郝宏伟,王睿,何哲,田人元,刘高强. 灵芝细胞中磷脂酸互作蛋白鉴定[J]. 生物工程学报, 2021, 37(9): 3293-3299

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2021-04-07
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2021-09-26
  • 出版日期:
您是第位访问者
生物工程学报 ® 2024 版权所有

通信地址:中国科学院微生物研究所    邮编:100101

电话:010-64807509   E-mail:cjb@im.ac.cn

技术支持:北京勤云科技发展有限公司