摘要
目的筛选人精子中与PHF7相互作用的蛋白质并分析其作用。方法构建质粒表达GST-PHF7融合蛋白,通过GST pull-down实验串联质谱检测人精子中与PHF7特异性结合的蛋白质,进行GO富集分析和KEGG通路分析。结果利用GST-PHF7诱饵蛋白从人精子细胞总蛋白中成功钓取靶蛋白。经过质谱检测,筛选出人精子中与PHF7特异结合的蛋白75个。GO分析结果表明,这些蛋白在生物学过程上主要富集在蛋白运输和细胞内定位等;在细胞成分上主要富集在细胞器,囊泡和外泌体等;在分子功能上主要富集在结合蛋白质,核酸和酶类等。KEGG分析结果表明,这些蛋白富集在细胞代谢和遗传信息的处理等通路。结论 PHF7与精子内多种蛋白质相互作用,调节蛋白合成,运输和降解及能量代谢等可能是其调节精子发生的机制。
Objective To screen proteins interact with PHF7 in response to male infertility and analyze the protein-protein interaction network. Methods GST-PHF7 fusion protein was expressed and purified for the next GST pull-down assay. The interacting proteins of PHF7 were detected by LC-MS/MS and then analyzed by GO enrichment and KEGG pathway databases. Results GST-PHF7 bait protein was successfully used to catch targets from the total proteins of human sperms and 75 interacting proteins were annotated. These proteins are involved in intracellular protein transport,cellular localization etc. in biological process;intracellular vesicle, cytoplasm, vesicle etc. in cellular component;protein binding, RNA binding, enzyme binding etc. in molecule function and intracellular protein transport, cellular localization,drug metabolic process, catabolic process etc. in KEGG pathways. Conclusions PHF7 and its interacting proteins may play a role in spermatogenesis by regulating protein transport and energy metabolic process.
作者
曹梦阳
林浩成
姜辉
Cao Mengyang;Lin Haocheng;Jiang Hui(Department of Urology,Peking University Third Hospital,Beijing 100191,China;Reproductive Medicine Center,Peking University Third Hospital,Beijing 100191,China;Department of Andrology,Peking University Third Hospital,Beijing 100191,China;Department of Human Sperm Bank,Peking University Third Hospital,Beijing 100191,China)
出处
《中国男科学杂志》
CAS
CSCD
2020年第2期15-22,共8页
Chinese Journal of Andrology
