摘要
在去除母乳中高丰度蛋白的基础上,运用蛋白质组学技术分析母乳中具有重要生物学特定功能的低丰度蛋白。以1月左右的成熟乳为实验材料,先通过免疫亲和层析和ProteoMiner低丰度蛋白富集试剂盒2种前处理方法去除高丰度蛋白,再进行蛋白质组学分析。结果表明,2种方法都有去除高丰度蛋白的效果,免疫亲和层析法处理后鉴定出蛋白质49种,ProteoMiner低丰度蛋白富集试剂盒处理后鉴定出蛋白质172种。对合计鉴定出的186种蛋白进行生物信息学分析,Gene Ontology(GO)注释分析发现,这些蛋白质功能主要集中在细胞部分、细胞过程和结合活性等方面。以Pathway数据库作为参考,依据代谢途径可将蛋白质定位到201个代谢途径分支,涉及PI3K-Akt信号通路、磷脂酶D信号通路等。与Cluster of Orthologous Groups of proteins(COG)数据库进行比对表明,这些蛋白质主要参与碳水化合物的运输和新陈代谢、信号转导机制等生命活动。研究结果不仅有助于了解母乳低丰度蛋白的功能,而且可为婴幼儿的健康成长和疾病预防提供理论依据,为母乳喂养的保护作用机制及有关乳腺疾病研究提供技术支撑。
After removing high-abundance proteins from breast milk,we applied proteomics to analyze low-abundance proteins with special important biofunctions in breast milk.Mature milk about 1 month old was used for this study,and high-abundance proteins were removed by immunoaffinity chromatography or ProteoMiner low-abundance protein enrichment kit,followed by proteomic analysis.The results showed that both methods were effective in removing highabundance proteins.A total of 186 low-abundance proteins were identified by the two methods,49 proteins of which were identified by immunoaffinity chromatography,and 172 by ProteoMiner low-abundance protein enrichment kit.According to bioinformatics analysis and gene ontology(GO)functional annotation analysis,the functions of the identified proteins mainly included serving as cellular components,participating in biological processes in cells and exerting binding activities.Using the Pathway database as a reference,these proteins were found to be involved in 201 metabolic pathway branches,including the PI3K-Akt signaling pathway and the phospholipase D signaling pathway.Compared with the Cluster of Orthologous Groups of proteins(COG)database,these proteins mainly participated in such biological activities as carbohydrate transport and metabolism,and signal transduction mechanisms.These results not only help to understand the functions of low-abundance proteins in breast milk,but also provide a theoretical basis for healthy growth and disease prevention in infants and young children as well as provide technical support for research on the protective mechanism of breastfeeding and on breast diseases.
作者
靳登鹏
周雄
刘焕
杨娇
黄卓权
卜玲玲
柯倩华
柳春红
JIN Dengpeng;ZHOU Xiong;LIU Huan;YANG Jiao;HUANG Zhuoquan;BU Lingling;KE Qianhua;LIU Chunhong(Guangdong Provincial Key Laboratory of Food Quality and Safety,Laboratory of Quality&Safety Risk Assessment for Agro-products on Storage and Preservation(Guangzhou),Ministry of Agriculture and Rural Affairs,College of Food Science,South China Agricultural University,Guangzhou 510642,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2020年第24期54-60,共7页
Food Science
基金
“十三五”国家重点研发计划重点专项(2017YFD0400602)。
关键词
母乳
多克隆抗体
免疫亲和层析
蛋白质组
breast milk
polyclonal antibody
immunoaffinity chromatography
proteome
