Proteomic analysis reveals proteins and pathways associated with lactation in bovine mammary epithelial cell-derived exosomes
Proteomic analysis reveals proteins and pathways associated with lactation in bovine mammary epithelial cell-derived exosomes
复制标题
蛋白质组学分析揭示了牛乳腺上皮细胞衍生的外泌体中与泌乳相关的蛋白质和途径
DOI：
10.1021/acs.jproteome.0c00176
复制
发表时间：
2020
影响因子：
4.4
通讯作者：
Gao Xuejun
中科院分区：
文献类型：
--
作者：
Zhang Minghui;Ma Zonghua;Li Rui;Guo Siqi;Qiu Youwen;Gao Xuejun
乳源性外泌体已被报道，它参与了许多生物过程。来自乳腺的外泌体尚不清楚，它们与乳腺泌乳的关系以及乳源性外泌体的起源在很大程度上尚不清楚。本研究旨在研究牛乳腺上皮细胞（BMECs）外泌体的蛋白质组，并将其与数据库中的乳源外泌体进行比较。采用联合超离心方法将bmec来源的外泌体从bmec培养上清中分离出来，并通过western blot分析鉴定外泌体的纯度。液相色谱串联质谱法鉴定了bmec衍生外泌体中的638个蛋白。MS数据存储到公共存储库ProteomeXchange，数据集标识符：https://www.iprox.org/page/PSV023.html;?url=1590961453176tKpa。基因本体注释和京都基因与基因组百科（KEGG）通路分析表明，这些蛋白与特定的生物学过程和代谢分子功能相关。将这些蛋白与乳源外泌体蛋白数据库进行交叉比较，发现77个共同表达蛋白（cep）存在于BMEC和乳源外泌体中。对这些cep的KEGG通路分析表明，它们主要参与bmec中与乳生物合成相关的信号通路。在这些cep中，有6种蛋白与泌乳功能有关。western blot分析发现，甲硫氨酸和β-雌二醇刺激bmec后，bmec衍生外泌体中这6种蛋白的表达增加。综上所述，bmec衍生外泌体的蛋白质组显示它们与bmec中的牛奶生物合成有关，可能是牛奶衍生外泌体的来源。
Milk-derived exosomes have been reported, which are involved in many biological processes. The exosomes derived from mammary glands are not known yet, and their relationship with mammary gland lactation and the origin of milk-derived exosomes are largely unclear. The present study aimed to investigate the proteome of exosomes derived from bovine mammary epithelial cells (BMECs) and compare them with milk-derived exosomes in the database. BMEC-derived exosomes were successfully separated from the culture supernatant of BMECs by a combined ultracentrifugation approach, and the purity of exosomes was identified by western blot analysis. Liquid chromatography with tandem mass spectrometry identified 638 proteins in BMEC-derived exosomes. The MS data were deposited into the PUBLIC repository ProteomeXchange, dataset identifier(s): https://www.iprox.org/page/PSV023.html;?url=1590961453176tKpa. Gene Ontology annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that these proteins were associated with specific biological processes and molecular functions of metabolism. Cross comparison of these proteins with the protein database of milk exosomes showed that 77 common expressed proteins (CEPs) were in both BMEC- and milk-derived exosomes. The KEGG pathway analysis for these CEPs showed that they were mainly involved in signaling pathways associated with milk biosynthesis in BMECs. Among these CEPs, six proteins have been previously reported to be associated with the lactation function. The western blot analysis detected that expression of these six proteins in BMEC-derived exosomes was increased after the stimulation of methionine and β-estradiol on BMECs. In summary, the proteome of BMEC-derived exosomes reveals that they are associated with milk biosynthesis in BMECs and might be a source of milk-derived exosomes.