摘要
目的利用iBAQ(intensity-based absolute-protein-quantification)非标记定量蛋白质组学分析鉴定生物被膜形成能力不同的鲍曼不动杆菌株蛋白质组学差异。方法64株临床分离的鲍曼不动杆菌株按生物被膜形成能力不同分为强组和弱组,收集两组菌株蛋白质样本进行FASP(filter-aided sample preparation)酶解,酶解产物进行LC-MS/MS质谱分析。使用Maxquant软件对LC-MS/MS原始文件进行查库以及iBAQ非标定量分析。Maxquant软件查库文件使用Perseus软件分析,通过GO(Gene Ontology)数据库对鉴定出的差异蛋白及特异蛋白从生物学过程、细胞成分和分子功能3方面进行功能注释,通过KAAS(KEGG automatic annotation server)分析差异蛋白涉及的代谢通路。结果实验通过LC-MS/MS分析共鉴定到1120个肽段,457个蛋白质,其中13个蛋白质的表达在生物被膜形成能力强组和弱组间差异具有统计学意义,其中7个蛋白质在弱组显著性低表达,6个在强组显著性低表达。另外,还鉴定出在生物被膜形成能力弱组(7个)和强组(5个)特异性表达的蛋白质,并对其参与的细胞过程及信号通路进行了分析。结论基于iBAQ的非标记定量蛋白质组学分析方法鉴定出多种与生物被膜形成能力相关的蛋白质,可为分析生物被膜形成的原因、机制及治疗提供参考。
Objective To analyze the proteomic differences of biofilm with different formation ability in Acinetobacter baumannii clinical isolates by using intensity based on absolute quantification(iBAQ)methods.Methods Biofilm models of 64 strains of Acinetobacter baumannii collected in our hospital were divided into strong biofilm-forming ability group and weak biofilm-forming ability group.Proteins were collected from these two groups respectively.After filter-aided sample preparation(FASP)protein digestion,the peptides were subjected to LC-MS/MS,and the obtained raw files were searched using Maxquant software and analyzed using Perseusn software.Gene Ontology(GO)terms were extracted for each peptide using the existing annotations.These GO terms were assigned to the biological process,molecular function and cellular compartments.KEGG automatic annotation server(KAAS)was used to analyze established pathway associations of differentially expressed proteins.Results In this study,a total of 1120 peptides and 457 proteins were identified between the two groups.Among all these 457 proteins,13 proteins were found to be significantly differentially expressed in these two groups,including 6 ones significantly lowly expressed in weak biofilm-forming ability groups and 7 significantly lowly expressed in strong biofilm-forming ability groups.We also identified 7 and 5 proteins specifically expressed in weak and strong biofilm-forming ability groups,respectively.Conclusion iBAQ method can identify diversified proteins associated with biofilm formation ability.These proteins provide reference for further research on biofilms.
作者
杨妮
雷莉
王海
邬媛
孙媛
张明
张正良
YANG Ni;LEI Li;WANG Hai;WU Yuan;SUN Yuan;ZHANG Ming;ZHANG Zheng-liang(Department of Emergency,The Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004,China)
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2020年第1期33-38,83,共7页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
陕西省卫生健康科研基金项目(No.2018D057)
陕西省中医药管理局科研项目(No.JCMS037)~~
